Kooperativer Bibliotheksverbund

UID:

Format: 1 online resource (XIII, 366 p. 74 illus., 62 illus. in color.)

Edition: 1st ed. 2021.

ISBN: 1-0716-1514-9

Series Statement: Methods in Molecular Biology, 2326

Content: This detailed book provides an accessible compendium of up-to-date methods in the fields of environmental toxicology, molecular toxicology, and toxicogenomics. Organized into four major sections, the volume examines methods utilizing model animal species, such as nematode, fruit fly, mice, chicken, and amphibians, methods using plants to study chemical toxicity, applying the Ames assay to chemical mutagenicity study, as well as methods for environmental chemical analysis. Although this book is divided into these parts, the methods can be used across species. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Environmental Toxicology and Toxicogenomics: Principles, Methods, and Applications serves as a valuable resource for the scientific community, particularly for young scientists and graduate and undergraduate students, inspiring more research in the vitally important field of environmental toxicity, molecular toxicology, and toxicogenomics.

Note: Detection of Caenorhabditis elegans Germ Cell Apoptosis Following Exposure to Environmental Contaminant Mixtures: A Crude Oil-Dispersant Mixture Example -- High-Throughput Measurement for Toxic Effects of Metal Mixtures in Caenorhabditis elegans -- Evaluations of Environmental Pollutants-Induced Mitochondrial Toxicity Using Caenorhabditis elegans as a Model System -- Methods to Assay the Behavior of Drosophila melanogaster for Toxicity Study -- Investigating the Joint Effects of Pesticides and Ultraviolet-B Radiation in Xenopus laevis and Other Amphibians -- Identification of Stable Reference Genes for Toxicogenomic and Gene Expression Analysis -- Semi-Quantitative RT-PCR: An Effective Method to Explore the Regulation of Gene Transcription Level Affected by Environmental Pollutants -- Employing Multiple New Neurobiological Methods to Investigate Environmental Neurotoxicology in Mice -- Analyses of Epigenetic Modification in Environmental Pollutants-Induced Neurotoxicity -- The Application of Omics Technologies in the Research of Neurotoxicology -- Flow Cytofluorometric Analysis of Molecular Mechanisms of Premature Red Blood Cell Death -- Practical Methods and Technologies in Environmental Epidemiology -- In Ovo Early-in-Life Inhalation Exposure to Gas/Aerosol with a Chicken Embryo Model -- DNA Damage in Liver Cells of the Tilapia Fish Oreochromis mossambicus Larva Induced by the Insecticide Cyantraniliprole at Sublethal Doses During Chronic Exposure -- Impact of Nanoparticles on Plant Growth, Development, and Biomass -- Biochemical and Physiological Toxicity of Nanoparticles in Plant -- Determination of Oxidative Stress and Antioxidant Enzyme Activity for Physiological Phenotyping During Heavy Metal Exposure -- Comprehensive Phytotoxicity Assessment Protocol for Engineered Nanomaterials -- Detection of Cadmium Toxicity in Plant -- Mutagenicity Evaluation of Nanoparticles by the Ames Assay -- Dispersive Solid Phase Extraction of Multiresidue Pesticides in Food and Water Samples -- Quantitative Analysis of Multiresidue Pesticides Using Gas Chromatography/Mass Spectrometry -- Determination of the N-Nitroso Compounds in Mouse Following RDX Exposure -- Determination of Metal Content in Drosophila melanogaster During Metal Exposure -- Microplastics: A Review of Methodology for Sampling and Characterizing Environmental and Biological Samples.

Additional Edition: ISBN 1-0716-1513-0

Language: English

Keywords: Llibres electrònics ; Llibres electrònics ; Llibres electrònics ; Llibres electrònics

DOI: 10.1007/978-1-0716-1514-0

UID:

Format: 1 online resource (X, 277 p. 55 illus., 28 illus. in color.)

Edition: 1st ed. 2013.

ISBN: 1-62703-212-6

Series Statement: Methods in Molecular Biology, 958

Content: Cotton is the most important textile and cash crop and is widely cultivated in more than 70 countries, including the United States, China, and India. Because of its long life cycle and complicated genetic background, it is hard to improve cotton using traditional breeding techniques although it has made much progress in the last several decades. Currently, transgenic techniques have become a powerful tool to improve cotton, and transgenic cotton is among the first commercially genetically modified crops. Transgenic Cotton: Methods and Protocols provides a comprehensive collection of methods for creating and monitoring transgenic cotton and its application on agricultural and basic research. Divided into five convenient sections, topics covered include the current status and perspectives of transgenic cotton, the principle and methods for making transgenic cotton, the methods for detecting foreign gene copy and expression in transgenic plants, the improvement of cotton using transgenic technology, and finally the methods for monitoring the potential impact of transgenic cotton on the environment, including gene flow. Written in the successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.   Authoritative and easily accessible, Transgenic Cotton: Methods and Protocols will serve as an excellent resource for scientists as well as graduate students who work on transgenic plants, plant genetics, molecular biology and agricultural sciences.

Note: Bibliographic Level Mode of Issuance: Monograph , Transgenic Cotton: From Biotransformation Methods to Agricultural Application -- Genetically Modified Cotton in India and Detection Strategies -- Agrobacterium-Mediated Transformation of Cotton -- Biolistic Transformation of Cotton Zygotic Embryo Meristem -- Biolistic Transformation of Cotton Embryogenic Cell Suspension Cultures -- Cotton Transformation via Pollen Tube Pathway -- Silicon Carbide Whisker-Mediated Transformation of Cotton (Gossypium hirsutum L.) -- Investigating Transgene Integration and Organization in Cotton (Gossypium hirsutum L.) Genome -- Estimating the Copy Number of Transgenes in Transformed Cotton by Real-Time Quantitative PCR -- Development of Enzyme-Linked Immunosorbant (ELISA) Assay for the Detection of Bt Protein in Transgenic Cotton -- DNA-Based Diagnostics for Genetically Modified Cotton: Decaplex PCR Assay to Differentiate MON531 and MON15985 Bt Cotton Events -- A Simple and Rapid Method for Determining Transgenic Cotton Plants -- An Efficient Grafting Technique for Recovery of Transgenic Cotton Plants -- Inheritance of Transgenes in Transgenic Bt Lines Resistance to Helicoerpa armigera in Upland Cotton -- Agrobacterium rhizogenes-Induced Cotton Hairy Root Culture as an Alternative Tool for Cotton Functional Genomics -- Overexpression of miR 156 in Cotton via Agrobacterium-Mediated Transformation -- Development of Transgenic CryIA(c)+GNA Cotton Plants via Pollen Tube Pathway Method Confers Resistance to Helicoverpa armigera and Aphis gossypii Glover -- Agrobacterium-Mediated Transformation of Cotton (Gossypium hirsutum) Shoot Apex with a Fungal Phytase Gene Improves Phosphorus Acquisition -- Genetic Transformation of Cotton with a Harpin-Encoding Gene hpaXoo Confers an Enhanced Defense Response Against Verticillium dahliae Kleb -- Development of Insect-Resistant Transgenic Cotton with Chimeric TVip3A* Accumulating in Chloroplasts -- Determining Gene Flow in Transgenic Cotton. , English

Additional Edition: ISBN 1-62703-211-8

Language: English

DOI: 10.1007/978-1-62703-212-4

UID:

Format: 1 online resource (XI, 240 p.)

Edition: 1st ed. 2010.

ISBN: 1-60761-768-4 , 1-60761-769-2

Series Statement: Methods in Molecular Biology, 650

Content: Recent stem cell research has revealed that miRNA and RNAi-mediated gene regulation is one of the vital determinates controlling the state of cell differentiation, with the small RNAs serving as key elements involved in regulatory network control of pluripotent cell fate determination. In RNAi and microRNA-Mediated Gene Regulation in Stem Cells: Methods, Protocols, and Applications, expert authors from laboratories across the globe contribute an accessible compendium of up-to-date, proven methods focused on the study of the titular topic. Divided into three sections, the book first gives a brief introduction to RNAi and miRNAs in stem cells, with a focus on the current status of research and future perspectives, then it continues with detailed methods and protocols for RNAi screening, transfection, and the knockdown of specific genes and pathways in several animal species, including humans and mice, concluding with a section on recently developed methods for identification of miRNAs, including a general protocol for preparation and analysis of miRNA libraries for deep sequencing, knock down of a specific gene using miRNA-based shRNA, and miRNA expression analysis using qRT-PCR. Written in the highly successful Methods in Molecular Biology™ series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes highlighting tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, RNAi and microRNA-Mediated Gene Regulation in Stem Cells: Methods, Protocols, and Applications serves as a valuable resource for scientists and aspiring graduate students interested in the intersection of RNAi, miRNA, and stem cell molecular biology and the exciting areas of medicine, including regenerative medicine, aging, cancer, and neurological disorders, that can be advanced through this expanding area of research.

Note: Bibliographic Level Mode of Issuance: Monograph , RNAi in Stem Cells: Current Status and Future Perspectives -- A Brief Introduction to RNAi and MicroRNAs in Stem Cells -- RNA Interference -- Forward RNAi Screens in Human Stem Cells -- A Recessive Genetic Screen for Components of the RNA Interference Pathway in Mouse Embryonic Stem Cells -- Construction and Application of Random dsRNA Interference Library for Functional Genetic Screens in Embryonic Stem Cells -- Establishing Efficient siRNA Knockdown in Stem Cells Using Fluorescent Oligonucleotides -- Loss-of-Function Studies in Mouse Embryonic Stem Cells Using the pHYPER shRNA Plasmid Vector -- Regulation and/or Repression of Cholinergic Differentiation of Murine Embryonic Stem Cells Using RNAi Directed Against Transcription Factor L3/Lhx8 -- Silencing of Rho-GDI? by RNAi Promotes the Differentiation of Neural Stem Cells -- RNAi Knockdown of Redox Signaling Protein Ape1 in the Differentiation of Mouse Embryonic Stem Cells -- Proteins Involved in Cell Migration from Glioblastoma Neurospheres Analyzed by Overexpression and siRNA-Mediated Knock-Down -- An Efficient Transfection Method for Mouse Embryonic Stem Cells -- Semi-quantitative Analysis of Transient Single-Cell Gene Expression in Embryonic Stem Cells by Femtoinjection -- MicroRNAs -- Preparation and Analysis of MicroRNA Libraries Using the Illumina Massively Parallel Sequencing Technology -- Assessing In Vivo MicroRNA Function in the Germline Stem Cells of the Drosophila Ovary -- Monitoring MicroRNA Expression During Embryonic Stem-Cell Differentiation Using Quantitative Real-Time PCR (qRT-PCR) -- Engineering Human Mesenchymal Stem Cells to Release Adenosine Using miRNA Technology -- Efficient Gene Knockdowns in Mouse Embryonic Stem Cells Using MicroRNA-Based shRNAs. , English

Additional Edition: Printed edition: ISBN 9781607617686

Language: English

DOI: 10.1007/978-1-60761-769-3

UID:

Format: 1 online resource (554 pages)

Edition: First edition.

ISBN: 0-443-13230-5

Note: Front cover -- Half title -- Title -- Copyright -- Contents -- Contributors -- Editor's Biography -- Series Preface -- Preface -- Section I The CRISPR/Cas toolkit for horticultural crops -- Chapter 1 CRISPRized fruit, vegetable, and ornamental crops: A note from editors -- 1.1 Introduction -- 1.2 What are genome-editing techniques? -- 1.2.1 Meganucleases -- 1.2.2 Zinc-finger nucleases -- 1.2.3 Transcriptional activator-like effector nucleases -- 1.2.4 CRISPR-Cas systems -- 1.3 CRISPR: a revolutionary technique for genetic improvement of plants -- 1.3.1 Hybrid breeding -- 1.3.2 Improving crop quality -- 1.3.3 Improving abiotic stress -- 1.3.4 Improve biotic stress tolerance -- 1.4 Accelerating horticultural plant domestication -- 1.5 Challenges -- 1.6 Conclusion -- References -- Chapter 2 Evolution of genome editing technologies -- 2.1 Introduction -- 2.2 DNA repairs system: a key in developing genome editing tools -- 2.3 The rise of the genome editing era -- 2.3.1 Homologous recombination -- 2.3.2 Meganucleases -- 2.3.3 Zinc-finger nucleases -- 2.3.4 TALENs -- 2.3.5 CRISPR-Cas9 -- 2.4 Advances in genome editing era -- 2.4.1 Base editors -- 2.4.2 Prime editors -- 2.5 Applications of gene editing technologies in agriculture -- 2.6 Application of gene editing technologies in horticulture crops -- 2.6.1 Enhance resistance to stresses -- 2.6.2 Improve fruit quality -- 2.6.3 Modify cultivation traits -- 2.6.4 Off-target analysis -- 2.7 Conclusion and future prospect -- References -- Chapter 3 CRISPR-Cas: Effectors, mechanism, and classification -- 3.1 Introduction -- 3.2 Objectives -- 3.3 CRISPR-Cas9 -- from adoptive immunity to gene editing tool -- 3.4 CRISPR-Cas9 system structure -- 3.5 CRISPR-Cas system working principle -- 3.6 Approaches for the classification of CRISPR-Cas system -- 3.7 Classification of CRISPR-Cas systems. , 3.7.1 CRISPR-Cas classes -- 3.7.2 Class 1 systems -- 3.7.3 Class 2 systems -- 3.8 Distribution of CRISPR-Cas system -- 3.9 Core and ancillary Cas genes -- 3.10 Origin and evolution of CRISPR-Cas9 -- 3.11 Anti-CRISPR defense -- 3.12 Conclusion -- References -- Chapter 4 Bioinformatics tools and databases in genome editing for plants -- 4.1 Introduction -- 4.2 Mechanism of genome editing techniques -- 4.2.1 Zinc-finger nucleases -- 4.2.2 Transcription activator-like effector nucleases -- 4.2.3 CRISPR-Cas system -- 4.3 Importance of guide RNA in genome editing -- 4.3.1 Efficient gene annotation: a key to unlocking genetic potential -- 4.3.2 Application-specific guidelines -- 4.3.3 Data-driven optimal design -- 4.4 gRNA designing process: a brief overview -- 4.4.1 Strategies for selecting highly efficient gRNAs -- 4.4.2 Key considerations for genome-wide CRISPR library design -- 4.4.3 The power of specificity: CRISPR-Cas technology in precision gene editing -- 4.4.4 Factors affecting specificity -- 4.4.5 Targeted CRISPR applications: key gRNA design considerations -- 4.5 Bioinformatics tools for designing gRNA -- 4.6 gRNA design tools: plant-specific databases -- 4.6.1 CRISPR-P -- 4.6.2 CRISPOR -- 4.6.3 E-CRISP -- 4.6.4 sgRNAcas9 -- 4.6.5 CRISPR-ERA -- 4.6.6 Cas-OFFinder -- 4.6.7 PlantPAN 3.0 -- 4.6.8 CRISPR-PLANT 2.0 -- 4.6.9 CRISPR-GE -- 4.6.10 PlantCARE-SG -- 4.7 Challenges and future prospects -- 4.7.1 Ethical considerations -- 4.7.2 Regulatory challenges -- 4.7.3 Future directions -- 4.8 Conclusion -- References -- Chapter 5 CRISPR workflow solutions: Cargos and versatile delivery platforms in genome editing -- 5.1 Introduction -- 5.2 CRISPR delivery formats -- 5.2.1 DNA-based delivery -- 5.2.2 mRNA format -- 5.2.3 RNP format -- 5.3 Viral-mediated CRISPR delivery -- 5.3.1 Adeno-associated viral vectors -- 5.3.2 Lentiviruses. , 5.3.3 Adenoviral vectors -- 5.3.4 Bacteriophage -- 5.4 Nonviral and chemical based CRISPR delivery methods -- 5.4.1 Polymeric-based nanoparticles -- 5.4.2 Inorganic nanoparticles -- 5.5 Physical delivery methods -- 5.5.1 Mechanical transfection -- 5.5.2 Microinjection -- 5.5.3 Electroporation -- 5.5.4 Hydrodynamic injection -- 5.5.5 Microfluidics chips -- 5.5.6 Acoustoporation -- 5.5.7 Magneto-transfection -- 5.5.8 Laser optoporation -- 5.5.9 iTOP (induced transduction by osmocytosis and propanebetaine) -- 5.5.10 Filtroporation -- 5.6 Delivery of CRISPR-Cas systems in plants -- 5.6.1 Indirect methods -- 5.6.2 Direct method -- 5.7 Future outlooks in CRISPR-Cas system delivery -- References -- Chapter 6 CRISPR-based techniques and their application in plants -- 6.1 Introduction -- 6.2 Components and basic structure of the CRISPR-Cas system -- 6.3 CRISPR-Cas system classification -- 6.3.1 Class I CRISPR-Cas system -- 6.3.2 Class II CRISPR-Cas system -- 6.4 CRISPR-Cas system applications -- 6.4.1 Gene editing -- 6.4.2 Antimicrobial -- 6.4.3 Bacterial strain typing and tracing -- 6.5 Application of the CRISPR tool in plants -- 6.6 Plant genome editing -- 6.6.1 Editing bases in plants -- 6.6.2 Prime edition in plants -- 6.6.3 Construction of the CRISPR-Cas system for plant genome editing -- 6.6.4 Epigenetic regulation with CRISPR-Cas -- 6.7 Applications of the CRISPR-Cas system in plants -- 6.7.1 Medicinal plants -- 6.7.2 Immunity against phytopathogenic viruses -- 6.7.3 DNA viruses -- 6.7.4 RNA viruses -- 6.7.5 Resistance to other pathogens -- 6.8 Limitations of gene editing using CRISPR-Cas in plants -- 6.9 Conclusion and future prospects -- Acknowledgment -- References -- Chapter 7 Base editing and prime editing in horticulture crops: Potential applications, challenges, and prospects -- 7.1 Introduction -- 7.2 Base editor -- 7.2.1 Cytosine base editor. , 7.2.2 Adenine base editor -- 7.2.3 Dual base editing -- 7.2.4 RNA base editor -- 7.3 Prime editing -- 7.4 Edits for important agronomic characteristics -- 7.5 Challenges and future prospects -- 7.6 Conclusion -- References -- Chapter 8 Multiplex genome editing in plants through CRISPR-Cas -- 8.1 Introduction -- 8.2 CRISPR-Cas9: a powerful approach for multiplex gene editing -- 8.3 Strategies of multiplex gene editing: unlocking the power of precise genetic modifications -- 8.3.1 Individual expression cassettes for each gRNA -- 8.3.2 Csy-4-based excision to multiple gRNAs -- 8.3.3 crRNA based CRISPR-Cas12a System -- 8.3.4 Self-cleaving ribozyme flanked gRNAs -- 8.3.5 tRNA-dependent expression and dispensation of gRNAs by endogenous RNases -- 8.3.6 Co-transfection of Cas9 protein loaded with multiple guide RNAs: a DNA free multiplex gene editing -- 8.4 Multiplex gene editing: reshaping the crop genetic improvement -- 8.4.1 Genetic diversity and crop domestication using multiplex gene editing -- 8.4.2 Increasing crop yield potential using multiplex gene editing -- 8.4.3 Improving crop quality by multiplex gene editing -- 8.4.4 Enhancing crop resistance to abiotic and biotic stresses -- 8.5 Applications of multiplex gene editing -- 8.5.1 Redefining gene editing: knocking out multiplex genes simultanously -- 8.5.2 Reshaping genomic landscape: restructuring of chromosomal segments -- 8.5.3 Breaking the exon boundaries: exon exchange as a catalyst for genetic variations -- 8.5.4 Mastering gene regulation: switching genes on and off through gene activation and repression -- 8.5.5 Tackling off-target challenges: advance genome editing with Cas9 dimers to reduce off-target effects -- 8.5.6 Unlocking epigenetic potential: epigenetic modifications for cellular reprogramming -- 8.5.7 Synergizing molecular farming with multiplex genome editing. , 8.5.8 Customizing the genetic code: harnessing multiplex base alternations for diverse genome modifications -- 8.6 Tackling complexity: insights and challenges of multiplex genome editing -- 8.7 Conclusion and future prospects -- References -- Chapter 9 CRISPR-Cas technologies for food and nutritional security -- 9.1 Introduction -- 9.2 History of CRISPR -- 9.3 Development of biofortified crops employing CRISPR-Cas mechanism -- 9.3.1 Genome editing for vitamin A enrichment -- 9.3.2 Genome editing for vitamin E enrichment -- 9.3.3 Genome editing for Fe enrichment -- 9.3.4 Genome editing for zinc enrichment -- 9.4 A breakthrough in editing a plant genome -- 9.5 Expanding the CRISPR-Cas editing toolkit -- 9.6 Recent frontiers in the CRISPR-Cas system -- 9.6.1 Base editing -- 9.6.2 Prime editing -- 9.6.3 Epigenome editing -- 9.6.4 De novo domestication -- 9.6.5 Multiplex genome editing -- 9.7 CRISPR-Cas for abiotic stresses -- 9.8 Targeting structural and regulatory genes -- 9.9 Application of CRISPR-Cas against abiotic stresses -- 9.9.1 Drought stress tolerance -- 9.9.2 Temperature stress tolerance -- 9.9.3 Salinity stress tolerance -- 9.9.4 Herbicide stress tolerance -- 9.10 Stress resilience crops for improved agricultural yield -- 9.11 Challenges and future prospects -- 9.12 Conclusion -- References -- Section II CRISPR mediated genome editing in horticultural crops -- Chapter 10 CRISPR-Cas9 genome editing of crops: Food and nutritional security -- 10.1 Introduction -- 10.1.1 The CRISPR-Cas9 toolbox -- 10.2 Components of CRISPR-Cas9 toolbox -- 10.2.1 CRISPR-Cas9-the mechanism -- 10.3 Process of genome editing via CRISPR-Cas9 -- 10.3.1 Progress of CRISPR-Cas9 in crop development -- 10.4 Genomics of produced crops -- 10.4.1 Rice -- 10.4.2 Other crops -- 10.5 Human exposure and public acceptance of the CRISPR-Cas9 system. , 10.6 CRISPR-mediated genome modification for food security.

Additional Edition: ISBN 0-443-13229-1

Language: English

UID:

Format: 1 online resource (XII, 327 p. 63 illus., 34 illus. in color.)

Edition: 2nd ed. 2019.

ISBN: 1-4939-8952-9

Series Statement: Methods in Molecular Biology, 1902

Content: This second edition provides a comprehensive collection of the cutting-edge methods for creating and monitoring transgenic cotton and its application on agricultural and basic research. Chapters detail current status and perspectives of transgenic cotton, principle and methods for making transgenic cotton, creating gene knockout lines, foreign gene copy and expression in transgenic plants, improvements to cotton using transgenic technology, and monitoring the potential impact of transgenic cotton on environment. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Transgenic Cotton: Methods and Protocols 2nd aims to be a resource for scientists as well as graduate students who work on transgenic plants, plant genetics, molecular biology, and agricultural sciences.

Note: Transgenic Cotton: From Biotransformation Methods to Agricultural Application -- Agrobacterium-mediated Genetic Transformation of Cotton -- Biolistic Transformation of Cotton Zygotic Embryo Meristem -- Novel Pollen Magnetofection System for Transformation of Cotton Plant with Magnetic Nanoparticles as Gene Carriers -- Biolistic Transformation of Cotton Embryogenic Cell Suspension Cultures -- Pollen Tube Pathway-Mediated Cotton Transformation -- Embryogenic Calli Explants and Silicon Carbide Whisker-mediated Transformation of Cotton (Gossypium hirsutum L.) -- Genome Editing in Cotton Using CRISPR/Cas9 System -- Tobacco Rattle Virus Induced Gene Silencing in Cotton -- Investigating Transgene Integration and Organization in Cotton (Gossypium hirsutum L.) Genome -- Estimating the Copy Number of Transgenes in Transformed Cotton by Real-time Quantitative PCR -- Development of an Enzyme-linked Immunosorbant (ELISA) Assay for the Detection of GM Protein in GM Crops/Produce -- Screening of Transgenic Cotton Based on a Porous Silicon Biosensor -- YC3.60-based Imaging Analysis on Calcium Level in Cotton Cells -- A Simple and Rapid Method for Determining Transgenic Cotton Plants Using a Marker Gene -- A Grafting Technique for Efficiently Transplanting Transgenic Regenerated Plants of Cotton -- Inheritance of Transgenes in Transgenic Bt lines Resistance to Helicoerpa armigera in Upland Cotton -- Cotton Hairy Root Culture as an Alternative Tool for Cotton Functional Genomics -- Overexpression of miRNA in Cotton via Agrobacterium-mediated Transformation -- Development of Transgenic CryIA(c)+GNA Cotton Plants via Pollen Tube Pathway Method Confers Resistance to Helicoverpa armigera and Aphis gossypii Glover -- Next Generation Transgenic Cotton: Pyramiding RNAi with Bt Counters Insect Resistance -- Genetic Transformation of Cotton with the Harpin-encoding Gene hpaXoo of Xanthomonas oryzae pv. Oryzae and Evaluation of Resistance against Verticillium Wilt -- Development of Insect-resistant Transgenic Cotton with Chimeric TVip3A* Accumulating in Chloroplasts -- Development of Virus Resistance Transgenic Cotton using Cotton Leaf Curl Virus Antisense ßC1 Gene -- Determining Pollen-mediated Gene Flow in Transgenic Cotton.

Additional Edition: ISBN 1-4939-8951-0

Language: English

DOI: 10.1007/978-1-4939-8952-2

UID:

Format: 1 online resource (XI, 311 p. 50 illus., 32 illus. in color.)

Edition: 1st ed. 2017.

ISBN: 1-4939-7108-5

Series Statement: Methods in Molecular Biology, 1622

Content: This detailed collection provides an accessible compendium of up-to-date methods focused on the study of RNAi and small regulatory miRNAs in stem cells. Beginning with a brief introductory section, the volume continues by exploring methods and protocols for RNAi screening, transfection, and the knockdown of specific genes and pathways in several animal species, including humans and mice, recently developed methods for miRNA expression and functional analysis, as well as usage of CRISPR/Cas 9 to knockout an individual gene for functional studies. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, list of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Practical and authoritative, RNAi and Small Regulatory RNAs in Stem Cells: Methods and Protocols aims to accelerate progress in this crucial field by reducing the time required to decipher and put into practice procedures published in the literature.

Note: RNAi Technique in Stem Cell Research: Current Status and Future Perspectives -- RNAi and MicroRNA-Mediated Gene Regulation in Stem Cells -- Forward RNAi Screens in Human Hematopoietic Stem Cells -- Silencing of ATP11B by RNAi-Induced Changes in Neural Stem Cell Morphology -- High-Efficiency Transfection of Glioblastoma Cells and a Simple Spheroid Migration Assay -- Construction and Application of Random dsRNA Interference Library for Functional Genetic Screens in Embryonic Stem Cells -- Efficient Depletion of Essential Gene Products for Loss-of-Function Studies in Embryonic Stem Cells -- Regulation and/or Repression of Cholinergic Differentiation of Murine Embryonic Stem Cells Using RNAi Directed Against Transcription Factor L3/Lhx8 -- A Recessive Genetic Screen for Components of the RNA Interference Pathway Performed in Mouse Embryonic Stem Cells -- RNAi Knockdown of Ape1 Gene in the Differentiation of Mouse Embryonic Stem Cells -- An Efficient Transfection Method for Differentiation and Cell Proliferation of Mouse Embryonic Stem Cells -- Implanting Glioblastoma Spheroids into Rat Brains and Monitoring Tumor Growth by MRI Volumetry -- RNAi-Based Techniques for the Analysis of Gene Function in Drosophila Germline Stem Cells -- In Vivo RNAi in the Drosophila Follicular Epithelium: Analysis of Stem Cell Maintenance, Proliferation, and Differentiation -- A Phenotype-Based RNAi Screening for Ras-ERK/MAPK Signaling-Associated Stem Cell Regulators in C. elegans -- Engineering Human Mesenchymal Stem Cells to Release Adenosine Using miRNA Technology -- Efficient Gene Knockdowns in Mouse Embryonic Stem Cells Using MicroRNA-Based shRNAs -- Using Quantitative Real-Time PCR to Detect MicroRNA Expression Profile During Embryonic Stem Cell Differentiation -- Genetic Tools for Self-Organizing Culture of Mouse Embryonic Stem Cells Via Small Regulatory RNA-Mediated Technologies, CRISPR/Cas9 and Inducible RNAi -- CRISPR-Cas9-Mediated Gene Editing in Mouse Spermatogonial Stem Cells.

Additional Edition: ISBN 1-4939-7106-9

Language: English

DOI: 10.1007/978-1-4939-7108-4

UID:

Format: XI, 261 Seiten , Ill., graph. Darst.

ISBN: 9781607617686

Series Statement: Springer protocols handbooks 650

Note: Text engl.

Language: English

Keywords: Stammzelle ; RNS-Interferenz ; Small RNA ; Genregulation ; Methode

UID:

Format: 1 online resource (554 pages)

Edition: First edition.

ISBN: 0-443-13230-5

Note: Front cover -- Half title -- Title -- Copyright -- Contents -- Contributors -- Editor's Biography -- Series Preface -- Preface -- Section I The CRISPR/Cas toolkit for horticultural crops -- Chapter 1 CRISPRized fruit, vegetable, and ornamental crops: A note from editors -- 1.1 Introduction -- 1.2 What are genome-editing techniques? -- 1.2.1 Meganucleases -- 1.2.2 Zinc-finger nucleases -- 1.2.3 Transcriptional activator-like effector nucleases -- 1.2.4 CRISPR-Cas systems -- 1.3 CRISPR: a revolutionary technique for genetic improvement of plants -- 1.3.1 Hybrid breeding -- 1.3.2 Improving crop quality -- 1.3.3 Improving abiotic stress -- 1.3.4 Improve biotic stress tolerance -- 1.4 Accelerating horticultural plant domestication -- 1.5 Challenges -- 1.6 Conclusion -- References -- Chapter 2 Evolution of genome editing technologies -- 2.1 Introduction -- 2.2 DNA repairs system: a key in developing genome editing tools -- 2.3 The rise of the genome editing era -- 2.3.1 Homologous recombination -- 2.3.2 Meganucleases -- 2.3.3 Zinc-finger nucleases -- 2.3.4 TALENs -- 2.3.5 CRISPR-Cas9 -- 2.4 Advances in genome editing era -- 2.4.1 Base editors -- 2.4.2 Prime editors -- 2.5 Applications of gene editing technologies in agriculture -- 2.6 Application of gene editing technologies in horticulture crops -- 2.6.1 Enhance resistance to stresses -- 2.6.2 Improve fruit quality -- 2.6.3 Modify cultivation traits -- 2.6.4 Off-target analysis -- 2.7 Conclusion and future prospect -- References -- Chapter 3 CRISPR-Cas: Effectors, mechanism, and classification -- 3.1 Introduction -- 3.2 Objectives -- 3.3 CRISPR-Cas9 -- from adoptive immunity to gene editing tool -- 3.4 CRISPR-Cas9 system structure -- 3.5 CRISPR-Cas system working principle -- 3.6 Approaches for the classification of CRISPR-Cas system -- 3.7 Classification of CRISPR-Cas systems. , 3.7.1 CRISPR-Cas classes -- 3.7.2 Class 1 systems -- 3.7.3 Class 2 systems -- 3.8 Distribution of CRISPR-Cas system -- 3.9 Core and ancillary Cas genes -- 3.10 Origin and evolution of CRISPR-Cas9 -- 3.11 Anti-CRISPR defense -- 3.12 Conclusion -- References -- Chapter 4 Bioinformatics tools and databases in genome editing for plants -- 4.1 Introduction -- 4.2 Mechanism of genome editing techniques -- 4.2.1 Zinc-finger nucleases -- 4.2.2 Transcription activator-like effector nucleases -- 4.2.3 CRISPR-Cas system -- 4.3 Importance of guide RNA in genome editing -- 4.3.1 Efficient gene annotation: a key to unlocking genetic potential -- 4.3.2 Application-specific guidelines -- 4.3.3 Data-driven optimal design -- 4.4 gRNA designing process: a brief overview -- 4.4.1 Strategies for selecting highly efficient gRNAs -- 4.4.2 Key considerations for genome-wide CRISPR library design -- 4.4.3 The power of specificity: CRISPR-Cas technology in precision gene editing -- 4.4.4 Factors affecting specificity -- 4.4.5 Targeted CRISPR applications: key gRNA design considerations -- 4.5 Bioinformatics tools for designing gRNA -- 4.6 gRNA design tools: plant-specific databases -- 4.6.1 CRISPR-P -- 4.6.2 CRISPOR -- 4.6.3 E-CRISP -- 4.6.4 sgRNAcas9 -- 4.6.5 CRISPR-ERA -- 4.6.6 Cas-OFFinder -- 4.6.7 PlantPAN 3.0 -- 4.6.8 CRISPR-PLANT 2.0 -- 4.6.9 CRISPR-GE -- 4.6.10 PlantCARE-SG -- 4.7 Challenges and future prospects -- 4.7.1 Ethical considerations -- 4.7.2 Regulatory challenges -- 4.7.3 Future directions -- 4.8 Conclusion -- References -- Chapter 5 CRISPR workflow solutions: Cargos and versatile delivery platforms in genome editing -- 5.1 Introduction -- 5.2 CRISPR delivery formats -- 5.2.1 DNA-based delivery -- 5.2.2 mRNA format -- 5.2.3 RNP format -- 5.3 Viral-mediated CRISPR delivery -- 5.3.1 Adeno-associated viral vectors -- 5.3.2 Lentiviruses. , 5.3.3 Adenoviral vectors -- 5.3.4 Bacteriophage -- 5.4 Nonviral and chemical based CRISPR delivery methods -- 5.4.1 Polymeric-based nanoparticles -- 5.4.2 Inorganic nanoparticles -- 5.5 Physical delivery methods -- 5.5.1 Mechanical transfection -- 5.5.2 Microinjection -- 5.5.3 Electroporation -- 5.5.4 Hydrodynamic injection -- 5.5.5 Microfluidics chips -- 5.5.6 Acoustoporation -- 5.5.7 Magneto-transfection -- 5.5.8 Laser optoporation -- 5.5.9 iTOP (induced transduction by osmocytosis and propanebetaine) -- 5.5.10 Filtroporation -- 5.6 Delivery of CRISPR-Cas systems in plants -- 5.6.1 Indirect methods -- 5.6.2 Direct method -- 5.7 Future outlooks in CRISPR-Cas system delivery -- References -- Chapter 6 CRISPR-based techniques and their application in plants -- 6.1 Introduction -- 6.2 Components and basic structure of the CRISPR-Cas system -- 6.3 CRISPR-Cas system classification -- 6.3.1 Class I CRISPR-Cas system -- 6.3.2 Class II CRISPR-Cas system -- 6.4 CRISPR-Cas system applications -- 6.4.1 Gene editing -- 6.4.2 Antimicrobial -- 6.4.3 Bacterial strain typing and tracing -- 6.5 Application of the CRISPR tool in plants -- 6.6 Plant genome editing -- 6.6.1 Editing bases in plants -- 6.6.2 Prime edition in plants -- 6.6.3 Construction of the CRISPR-Cas system for plant genome editing -- 6.6.4 Epigenetic regulation with CRISPR-Cas -- 6.7 Applications of the CRISPR-Cas system in plants -- 6.7.1 Medicinal plants -- 6.7.2 Immunity against phytopathogenic viruses -- 6.7.3 DNA viruses -- 6.7.4 RNA viruses -- 6.7.5 Resistance to other pathogens -- 6.8 Limitations of gene editing using CRISPR-Cas in plants -- 6.9 Conclusion and future prospects -- Acknowledgment -- References -- Chapter 7 Base editing and prime editing in horticulture crops: Potential applications, challenges, and prospects -- 7.1 Introduction -- 7.2 Base editor -- 7.2.1 Cytosine base editor. , 7.2.2 Adenine base editor -- 7.2.3 Dual base editing -- 7.2.4 RNA base editor -- 7.3 Prime editing -- 7.4 Edits for important agronomic characteristics -- 7.5 Challenges and future prospects -- 7.6 Conclusion -- References -- Chapter 8 Multiplex genome editing in plants through CRISPR-Cas -- 8.1 Introduction -- 8.2 CRISPR-Cas9: a powerful approach for multiplex gene editing -- 8.3 Strategies of multiplex gene editing: unlocking the power of precise genetic modifications -- 8.3.1 Individual expression cassettes for each gRNA -- 8.3.2 Csy-4-based excision to multiple gRNAs -- 8.3.3 crRNA based CRISPR-Cas12a System -- 8.3.4 Self-cleaving ribozyme flanked gRNAs -- 8.3.5 tRNA-dependent expression and dispensation of gRNAs by endogenous RNases -- 8.3.6 Co-transfection of Cas9 protein loaded with multiple guide RNAs: a DNA free multiplex gene editing -- 8.4 Multiplex gene editing: reshaping the crop genetic improvement -- 8.4.1 Genetic diversity and crop domestication using multiplex gene editing -- 8.4.2 Increasing crop yield potential using multiplex gene editing -- 8.4.3 Improving crop quality by multiplex gene editing -- 8.4.4 Enhancing crop resistance to abiotic and biotic stresses -- 8.5 Applications of multiplex gene editing -- 8.5.1 Redefining gene editing: knocking out multiplex genes simultanously -- 8.5.2 Reshaping genomic landscape: restructuring of chromosomal segments -- 8.5.3 Breaking the exon boundaries: exon exchange as a catalyst for genetic variations -- 8.5.4 Mastering gene regulation: switching genes on and off through gene activation and repression -- 8.5.5 Tackling off-target challenges: advance genome editing with Cas9 dimers to reduce off-target effects -- 8.5.6 Unlocking epigenetic potential: epigenetic modifications for cellular reprogramming -- 8.5.7 Synergizing molecular farming with multiplex genome editing. , 8.5.8 Customizing the genetic code: harnessing multiplex base alternations for diverse genome modifications -- 8.6 Tackling complexity: insights and challenges of multiplex genome editing -- 8.7 Conclusion and future prospects -- References -- Chapter 9 CRISPR-Cas technologies for food and nutritional security -- 9.1 Introduction -- 9.2 History of CRISPR -- 9.3 Development of biofortified crops employing CRISPR-Cas mechanism -- 9.3.1 Genome editing for vitamin A enrichment -- 9.3.2 Genome editing for vitamin E enrichment -- 9.3.3 Genome editing for Fe enrichment -- 9.3.4 Genome editing for zinc enrichment -- 9.4 A breakthrough in editing a plant genome -- 9.5 Expanding the CRISPR-Cas editing toolkit -- 9.6 Recent frontiers in the CRISPR-Cas system -- 9.6.1 Base editing -- 9.6.2 Prime editing -- 9.6.3 Epigenome editing -- 9.6.4 De novo domestication -- 9.6.5 Multiplex genome editing -- 9.7 CRISPR-Cas for abiotic stresses -- 9.8 Targeting structural and regulatory genes -- 9.9 Application of CRISPR-Cas against abiotic stresses -- 9.9.1 Drought stress tolerance -- 9.9.2 Temperature stress tolerance -- 9.9.3 Salinity stress tolerance -- 9.9.4 Herbicide stress tolerance -- 9.10 Stress resilience crops for improved agricultural yield -- 9.11 Challenges and future prospects -- 9.12 Conclusion -- References -- Section II CRISPR mediated genome editing in horticultural crops -- Chapter 10 CRISPR-Cas9 genome editing of crops: Food and nutritional security -- 10.1 Introduction -- 10.1.1 The CRISPR-Cas9 toolbox -- 10.2 Components of CRISPR-Cas9 toolbox -- 10.2.1 CRISPR-Cas9-the mechanism -- 10.3 Process of genome editing via CRISPR-Cas9 -- 10.3.1 Progress of CRISPR-Cas9 in crop development -- 10.4 Genomics of produced crops -- 10.4.1 Rice -- 10.4.2 Other crops -- 10.5 Human exposure and public acceptance of the CRISPR-Cas9 system. , 10.6 CRISPR-mediated genome modification for food security.

Additional Edition: ISBN 0-443-13229-1

Language: English

UID:

Format: 1 online resource (554 pages)

Edition: First edition.

ISBN: 0-443-13230-5

Note: Front cover -- Half title -- Title -- Copyright -- Contents -- Contributors -- Editor's Biography -- Series Preface -- Preface -- Section I The CRISPR/Cas toolkit for horticultural crops -- Chapter 1 CRISPRized fruit, vegetable, and ornamental crops: A note from editors -- 1.1 Introduction -- 1.2 What are genome-editing techniques? -- 1.2.1 Meganucleases -- 1.2.2 Zinc-finger nucleases -- 1.2.3 Transcriptional activator-like effector nucleases -- 1.2.4 CRISPR-Cas systems -- 1.3 CRISPR: a revolutionary technique for genetic improvement of plants -- 1.3.1 Hybrid breeding -- 1.3.2 Improving crop quality -- 1.3.3 Improving abiotic stress -- 1.3.4 Improve biotic stress tolerance -- 1.4 Accelerating horticultural plant domestication -- 1.5 Challenges -- 1.6 Conclusion -- References -- Chapter 2 Evolution of genome editing technologies -- 2.1 Introduction -- 2.2 DNA repairs system: a key in developing genome editing tools -- 2.3 The rise of the genome editing era -- 2.3.1 Homologous recombination -- 2.3.2 Meganucleases -- 2.3.3 Zinc-finger nucleases -- 2.3.4 TALENs -- 2.3.5 CRISPR-Cas9 -- 2.4 Advances in genome editing era -- 2.4.1 Base editors -- 2.4.2 Prime editors -- 2.5 Applications of gene editing technologies in agriculture -- 2.6 Application of gene editing technologies in horticulture crops -- 2.6.1 Enhance resistance to stresses -- 2.6.2 Improve fruit quality -- 2.6.3 Modify cultivation traits -- 2.6.4 Off-target analysis -- 2.7 Conclusion and future prospect -- References -- Chapter 3 CRISPR-Cas: Effectors, mechanism, and classification -- 3.1 Introduction -- 3.2 Objectives -- 3.3 CRISPR-Cas9 -- from adoptive immunity to gene editing tool -- 3.4 CRISPR-Cas9 system structure -- 3.5 CRISPR-Cas system working principle -- 3.6 Approaches for the classification of CRISPR-Cas system -- 3.7 Classification of CRISPR-Cas systems. , 3.7.1 CRISPR-Cas classes -- 3.7.2 Class 1 systems -- 3.7.3 Class 2 systems -- 3.8 Distribution of CRISPR-Cas system -- 3.9 Core and ancillary Cas genes -- 3.10 Origin and evolution of CRISPR-Cas9 -- 3.11 Anti-CRISPR defense -- 3.12 Conclusion -- References -- Chapter 4 Bioinformatics tools and databases in genome editing for plants -- 4.1 Introduction -- 4.2 Mechanism of genome editing techniques -- 4.2.1 Zinc-finger nucleases -- 4.2.2 Transcription activator-like effector nucleases -- 4.2.3 CRISPR-Cas system -- 4.3 Importance of guide RNA in genome editing -- 4.3.1 Efficient gene annotation: a key to unlocking genetic potential -- 4.3.2 Application-specific guidelines -- 4.3.3 Data-driven optimal design -- 4.4 gRNA designing process: a brief overview -- 4.4.1 Strategies for selecting highly efficient gRNAs -- 4.4.2 Key considerations for genome-wide CRISPR library design -- 4.4.3 The power of specificity: CRISPR-Cas technology in precision gene editing -- 4.4.4 Factors affecting specificity -- 4.4.5 Targeted CRISPR applications: key gRNA design considerations -- 4.5 Bioinformatics tools for designing gRNA -- 4.6 gRNA design tools: plant-specific databases -- 4.6.1 CRISPR-P -- 4.6.2 CRISPOR -- 4.6.3 E-CRISP -- 4.6.4 sgRNAcas9 -- 4.6.5 CRISPR-ERA -- 4.6.6 Cas-OFFinder -- 4.6.7 PlantPAN 3.0 -- 4.6.8 CRISPR-PLANT 2.0 -- 4.6.9 CRISPR-GE -- 4.6.10 PlantCARE-SG -- 4.7 Challenges and future prospects -- 4.7.1 Ethical considerations -- 4.7.2 Regulatory challenges -- 4.7.3 Future directions -- 4.8 Conclusion -- References -- Chapter 5 CRISPR workflow solutions: Cargos and versatile delivery platforms in genome editing -- 5.1 Introduction -- 5.2 CRISPR delivery formats -- 5.2.1 DNA-based delivery -- 5.2.2 mRNA format -- 5.2.3 RNP format -- 5.3 Viral-mediated CRISPR delivery -- 5.3.1 Adeno-associated viral vectors -- 5.3.2 Lentiviruses. , 5.3.3 Adenoviral vectors -- 5.3.4 Bacteriophage -- 5.4 Nonviral and chemical based CRISPR delivery methods -- 5.4.1 Polymeric-based nanoparticles -- 5.4.2 Inorganic nanoparticles -- 5.5 Physical delivery methods -- 5.5.1 Mechanical transfection -- 5.5.2 Microinjection -- 5.5.3 Electroporation -- 5.5.4 Hydrodynamic injection -- 5.5.5 Microfluidics chips -- 5.5.6 Acoustoporation -- 5.5.7 Magneto-transfection -- 5.5.8 Laser optoporation -- 5.5.9 iTOP (induced transduction by osmocytosis and propanebetaine) -- 5.5.10 Filtroporation -- 5.6 Delivery of CRISPR-Cas systems in plants -- 5.6.1 Indirect methods -- 5.6.2 Direct method -- 5.7 Future outlooks in CRISPR-Cas system delivery -- References -- Chapter 6 CRISPR-based techniques and their application in plants -- 6.1 Introduction -- 6.2 Components and basic structure of the CRISPR-Cas system -- 6.3 CRISPR-Cas system classification -- 6.3.1 Class I CRISPR-Cas system -- 6.3.2 Class II CRISPR-Cas system -- 6.4 CRISPR-Cas system applications -- 6.4.1 Gene editing -- 6.4.2 Antimicrobial -- 6.4.3 Bacterial strain typing and tracing -- 6.5 Application of the CRISPR tool in plants -- 6.6 Plant genome editing -- 6.6.1 Editing bases in plants -- 6.6.2 Prime edition in plants -- 6.6.3 Construction of the CRISPR-Cas system for plant genome editing -- 6.6.4 Epigenetic regulation with CRISPR-Cas -- 6.7 Applications of the CRISPR-Cas system in plants -- 6.7.1 Medicinal plants -- 6.7.2 Immunity against phytopathogenic viruses -- 6.7.3 DNA viruses -- 6.7.4 RNA viruses -- 6.7.5 Resistance to other pathogens -- 6.8 Limitations of gene editing using CRISPR-Cas in plants -- 6.9 Conclusion and future prospects -- Acknowledgment -- References -- Chapter 7 Base editing and prime editing in horticulture crops: Potential applications, challenges, and prospects -- 7.1 Introduction -- 7.2 Base editor -- 7.2.1 Cytosine base editor. , 7.2.2 Adenine base editor -- 7.2.3 Dual base editing -- 7.2.4 RNA base editor -- 7.3 Prime editing -- 7.4 Edits for important agronomic characteristics -- 7.5 Challenges and future prospects -- 7.6 Conclusion -- References -- Chapter 8 Multiplex genome editing in plants through CRISPR-Cas -- 8.1 Introduction -- 8.2 CRISPR-Cas9: a powerful approach for multiplex gene editing -- 8.3 Strategies of multiplex gene editing: unlocking the power of precise genetic modifications -- 8.3.1 Individual expression cassettes for each gRNA -- 8.3.2 Csy-4-based excision to multiple gRNAs -- 8.3.3 crRNA based CRISPR-Cas12a System -- 8.3.4 Self-cleaving ribozyme flanked gRNAs -- 8.3.5 tRNA-dependent expression and dispensation of gRNAs by endogenous RNases -- 8.3.6 Co-transfection of Cas9 protein loaded with multiple guide RNAs: a DNA free multiplex gene editing -- 8.4 Multiplex gene editing: reshaping the crop genetic improvement -- 8.4.1 Genetic diversity and crop domestication using multiplex gene editing -- 8.4.2 Increasing crop yield potential using multiplex gene editing -- 8.4.3 Improving crop quality by multiplex gene editing -- 8.4.4 Enhancing crop resistance to abiotic and biotic stresses -- 8.5 Applications of multiplex gene editing -- 8.5.1 Redefining gene editing: knocking out multiplex genes simultanously -- 8.5.2 Reshaping genomic landscape: restructuring of chromosomal segments -- 8.5.3 Breaking the exon boundaries: exon exchange as a catalyst for genetic variations -- 8.5.4 Mastering gene regulation: switching genes on and off through gene activation and repression -- 8.5.5 Tackling off-target challenges: advance genome editing with Cas9 dimers to reduce off-target effects -- 8.5.6 Unlocking epigenetic potential: epigenetic modifications for cellular reprogramming -- 8.5.7 Synergizing molecular farming with multiplex genome editing. , 8.5.8 Customizing the genetic code: harnessing multiplex base alternations for diverse genome modifications -- 8.6 Tackling complexity: insights and challenges of multiplex genome editing -- 8.7 Conclusion and future prospects -- References -- Chapter 9 CRISPR-Cas technologies for food and nutritional security -- 9.1 Introduction -- 9.2 History of CRISPR -- 9.3 Development of biofortified crops employing CRISPR-Cas mechanism -- 9.3.1 Genome editing for vitamin A enrichment -- 9.3.2 Genome editing for vitamin E enrichment -- 9.3.3 Genome editing for Fe enrichment -- 9.3.4 Genome editing for zinc enrichment -- 9.4 A breakthrough in editing a plant genome -- 9.5 Expanding the CRISPR-Cas editing toolkit -- 9.6 Recent frontiers in the CRISPR-Cas system -- 9.6.1 Base editing -- 9.6.2 Prime editing -- 9.6.3 Epigenome editing -- 9.6.4 De novo domestication -- 9.6.5 Multiplex genome editing -- 9.7 CRISPR-Cas for abiotic stresses -- 9.8 Targeting structural and regulatory genes -- 9.9 Application of CRISPR-Cas against abiotic stresses -- 9.9.1 Drought stress tolerance -- 9.9.2 Temperature stress tolerance -- 9.9.3 Salinity stress tolerance -- 9.9.4 Herbicide stress tolerance -- 9.10 Stress resilience crops for improved agricultural yield -- 9.11 Challenges and future prospects -- 9.12 Conclusion -- References -- Section II CRISPR mediated genome editing in horticultural crops -- Chapter 10 CRISPR-Cas9 genome editing of crops: Food and nutritional security -- 10.1 Introduction -- 10.1.1 The CRISPR-Cas9 toolbox -- 10.2 Components of CRISPR-Cas9 toolbox -- 10.2.1 CRISPR-Cas9-the mechanism -- 10.3 Process of genome editing via CRISPR-Cas9 -- 10.3.1 Progress of CRISPR-Cas9 in crop development -- 10.4 Genomics of produced crops -- 10.4.1 Rice -- 10.4.2 Other crops -- 10.5 Human exposure and public acceptance of the CRISPR-Cas9 system. , 10.6 CRISPR-mediated genome modification for food security.

Additional Edition: ISBN 0-443-13229-1

Language: English

UID:

Format: 1 online resource (XI, 311 p. 50 illus., 32 illus. in color.)

Edition: 1st ed. 2017.

ISBN: 1-4939-7108-5

Series Statement: Methods in Molecular Biology, 1622

Content: This detailed collection provides an accessible compendium of up-to-date methods focused on the study of RNAi and small regulatory miRNAs in stem cells. Beginning with a brief introductory section, the volume continues by exploring methods and protocols for RNAi screening, transfection, and the knockdown of specific genes and pathways in several animal species, including humans and mice, recently developed methods for miRNA expression and functional analysis, as well as usage of CRISPR/Cas 9 to knockout an individual gene for functional studies. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, list of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Practical and authoritative, RNAi and Small Regulatory RNAs in Stem Cells: Methods and Protocols aims to accelerate progress in this crucial field by reducing the time required to decipher and put into practice procedures published in the literature.

Note: RNAi Technique in Stem Cell Research: Current Status and Future Perspectives -- RNAi and MicroRNA-Mediated Gene Regulation in Stem Cells -- Forward RNAi Screens in Human Hematopoietic Stem Cells -- Silencing of ATP11B by RNAi-Induced Changes in Neural Stem Cell Morphology -- High-Efficiency Transfection of Glioblastoma Cells and a Simple Spheroid Migration Assay -- Construction and Application of Random dsRNA Interference Library for Functional Genetic Screens in Embryonic Stem Cells -- Efficient Depletion of Essential Gene Products for Loss-of-Function Studies in Embryonic Stem Cells -- Regulation and/or Repression of Cholinergic Differentiation of Murine Embryonic Stem Cells Using RNAi Directed Against Transcription Factor L3/Lhx8 -- A Recessive Genetic Screen for Components of the RNA Interference Pathway Performed in Mouse Embryonic Stem Cells -- RNAi Knockdown of Ape1 Gene in the Differentiation of Mouse Embryonic Stem Cells -- An Efficient Transfection Method for Differentiation and Cell Proliferation of Mouse Embryonic Stem Cells -- Implanting Glioblastoma Spheroids into Rat Brains and Monitoring Tumor Growth by MRI Volumetry -- RNAi-Based Techniques for the Analysis of Gene Function in Drosophila Germline Stem Cells -- In Vivo RNAi in the Drosophila Follicular Epithelium: Analysis of Stem Cell Maintenance, Proliferation, and Differentiation -- A Phenotype-Based RNAi Screening for Ras-ERK/MAPK Signaling-Associated Stem Cell Regulators in C. elegans -- Engineering Human Mesenchymal Stem Cells to Release Adenosine Using miRNA Technology -- Efficient Gene Knockdowns in Mouse Embryonic Stem Cells Using MicroRNA-Based shRNAs -- Using Quantitative Real-Time PCR to Detect MicroRNA Expression Profile During Embryonic Stem Cell Differentiation -- Genetic Tools for Self-Organizing Culture of Mouse Embryonic Stem Cells Via Small Regulatory RNA-Mediated Technologies, CRISPR/Cas9 and Inducible RNAi -- CRISPR-Cas9-Mediated Gene Editing in Mouse Spermatogonial Stem Cells.

Additional Edition: ISBN 1-4939-7106-9

Language: English

DOI: 10.1007/978-1-4939-7108-4