In:
Development, Growth & Differentiation, Wiley, Vol. 56, No. 1 ( 2014-01), p. 98-107
Abstract:
Genome editing using engineered nucleases such as transcription activator‐like effector nucleases ( TALEN s) has become a powerful technology for reverse genetics. In this study, we have described efficient detection methods for TALEN ‐induced mutations at endogenous loci and presented guidelines of TALEN design for efficient targeted mutagenesis in medaka, Oryzias latipes . We performed a heteroduplex mobility assay ( HMA ) using an automated microchip electrophoresis system, which is a simple and high‐throughput method for evaluation of in vivo activity of TALEN s and for genotyping mutant fish of F1 or later generations. We found that a specific pattern of mutations is dominant for TALEN s harboring several base pairs of homologous sequences in target sequence. Furthermore, we found that a 5′ T, upstream of each TALEN ‐binding sequence, is not essential for genomic DNA cleavage. Our findings provide information that expands the potential of TALEN s and other engineered nucleases as tools for targeted genome editing in a wide range of organisms, including medaka.
Type of Medium:
Online Resource
ISSN:
0012-1592
,
1440-169X
DOI:
10.1111/dgd.2014.56.issue-1
Language:
English
Publisher:
Wiley
Publication Date:
2014
detail.hit.zdb_id:
2020067-5
SSG:
12
