Methods for in vitro CRISPR/CasRx-Mediated RNA Editing

被引:18
作者
Chuang, Yu-Fan [1 ,2 ]
Wang, Peng-Yuan [1 ]
Kumar, Satheesh [2 ]
Lama, Suraj [2 ]
Lin, Fan-Li [1 ,2 ]
Liu, Guei-Sheung [2 ,3 ,4 ]
机构
[1] Chinese Acad Sci, Shenzhen Inst Adv Technol, Shenzhen Key Lab Biomimet Mat & Cellular Immunomo, Shenzhen, Peoples R China
[2] Univ Tasmania, Menzies Inst Med Res, Hobart, Tas, Australia
[3] Univ Melbourne, Dept Surg, Ophthalmol, East Melbourne, Vic, Australia
[4] Aier Eye Inst, Changsha, Peoples R China
基金
英国医学研究理事会;
关键词
CRISPR; Cas13d; CasRx; RNA editing; VEGF (Vascular endothelial growth factor); AAV (Adeno-associated virus);
D O I
10.3389/fcell.2021.667879
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
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).
引用
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页数:13
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