In:
Current Protocols in Nucleic Acid Chemistry, Wiley, Vol. 72, No. 1 ( 2018-03)
Abstract:
A synthetic 8‐mer, amphipathic, trans ‐acting poly‐2′‐ O ‐methyluridylic thiophosphate triester RNA element (2′‐OMeUtaPS) can be prepared using solid‐phase synthesis protocols. 2′‐OMeUtaPS efficiently mediates the delivery of uncharged polyA‐tailed phosphorodiamidate morpholino (PMO) sequences in HeLa pLuc 705 cells, as evidenced by flow cytometry measurements. In this cell line, 2′‐OMeUtaPS‐mediated transfection of an antisense polyA‐tailed PMO sequence induces alternative splicing of an aberrant luciferase pre‐mRNA splice site, leading to restoration of functional luciferase, as quantitatively measured using a typical luciferase assay. 2′‐OMeUtaPS is also potent at delivering an uncharged antisense polyA‐tailed PMO sequence in muscle cells of the mdx mouse model of muscular dystrophy; targeting the polyA‐tailed PMO sequence against a splice site of the pre‐mRNA encoding mutated dystrophin triggers an alternate splicing event that results in excision of the mutated exon (exon 23) from the pre‐mRNA and production of functional dystrophin, as demonstrated by agarose gel electrophoresis. © 2018 by John Wiley & Sons, Inc.
Type of Medium:
Online Resource
ISSN:
1934-9270
,
1934-9289
Language:
English
Publisher:
Wiley
Publication Date:
2018
detail.hit.zdb_id:
2179070-X
