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Ubiquitous and Tissue-specific RNA Targeting in Drosophila Melanogaster using CRISPR/CasRx

J Vis Exp. 2021-02; 
Ruichen Sun, Daniel Brogan, Anna Buchman, Ting Yang, Omar S Akbari
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Gene Synthesis … gRNAs (comprised of a 36-nt direct repeat [DR] and a 30-nt spacer) corresponding to different target genes followed by an extra DR and a seven-thymine terminator was synthesized and subcloned into the digested backbone using Gene Synthesis (GenScript USA, Inc.)?… Get A Quote

摘要

CasRx, a member of the RNA-targeting Cas13 family, is a promising new addition of the CRISPR/Cas technologies in efficient gene transcript reduction with an attractive off-target profile at both cellular and organismal levels. It is recently reported that the CRISPR/CasRx system can be used to achieve ubiquitous and tissue-specific gene transcript reduction in Drosophila melanogaster. This paper details the methods from the recent work, consisting of three parts: 1) ubiquitous?in vivo endogenous RNA targeting using a two-component CasRx system; 2) ubiquitous in vivo exogenous RNA targeting using a three-component CasRx system; and 3) tissue-specific in vivo RNA targeting using a three-component CasRx system. T... More

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