Methods for in vitro CRISPR/CasRx-Mediated RNA Editing

体外 CRISPR/CasRx 介导的 RNA 编辑方法

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作者:Yu-Fan Chuang, Peng-Yuan Wang, Satheesh Kumar, Suraj Lama, Fan-Li Lin, Guei-Sheung Liu

Abstract

Specific changes in the genome have been accomplished by the revolutionary gene-editing tool known as clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system. The advent of programmable RNA editing CRISPR/Cas nucleases has made this gene-editing tool safer and more precise. Specifically, CasRx, a family member of the Cas13d family, has shown great therapeutic potential. Here, we describe the in vitro methods of utilizing this powerful RNA editing platform and determine the RNA editing efficiencies for CasRx with different forms of guide RNAs (also known as gRNA or sgRNA).

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