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CRISPR/Cas9 with single guide RNA expression driven by small tRNA promoters showed reduced editing efficiency compared to a U6 promoter.
RNA ; 23(1): 1-5, 2017 01.
Artigo em Inglês | MEDLINE | ID: mdl-27742910
ABSTRACT
Multiplex genome engineering in vivo with CRISPR/Cas9 shows great promise as a potential therapeutic approach. The ability to incorporate multiple single guide RNA (sgRNA) cassettes together with Cas9 gene expression in one AAV vector could greatly enhance the efficiency. In a recent Method article, Mefferd and coworkers indicated that small tRNA promoters could be used to drive sgRNA expression to facilitate the construction of a more effective AAV vector. In contrast, we found that when targeting endogenous genomic loci, CRISPR/Cas9 with tRNA promoter-driven sgRNA expression showed much reduced genome editing activity, compared with significant cleavage with U6 promoter-driven sgRNA expression. Though the underlying mechanisms are still under investigation, our study suggests that the CRISPR/Cas9 system with tRNA promoter-driven sgRNA expression needs to be reevaluated before it can be used for therapeutic genome editing.
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Coleções: Bases de dados internacionais Base de dados: MEDLINE Assunto principal: RNA de Transferência / Regiões Promotoras Genéticas / RNA Guia / Exorribonucleases / Edição de Genes Idioma: Inglês Revista: RNA Assunto da revista: Biologia Molecular Ano de publicação: 2017 Tipo de documento: Artigo País de afiliação: China