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

TITLE=Methods for in vitro CRISPR/CasRx-Mediated RNA Editing

JOURNAL=Frontiers in Cell and Developmental Biology

VOLUME=Volume 9 - 2021

YEAR=2021

URL=https://www.frontiersin.org/journals/cell-and-developmental-biology/articles/10.3389/fcell.2021.667879

DOI=10.3389/fcell.2021.667879

ISSN=2296-634X

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 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).