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论文题目: CRISPR/Cas9 with single guide RNA expression driven by small tRNA promoters showed reduced editing efficiency compared to a U6 promoter
英文论文题目: CRISPR/Cas9 with single guide RNA expression driven by small tRNA promoters showed reduced editing efficiency compared to a U6 promoter
第一作者: Wei, YD; Qiu, Y; Chen, YH; Liu, GG; Zhang, YX; Xu, LW; Ding, QR
英文第一作者: Wei, YD; Qiu, Y; Chen, YH; Liu, GG; Zhang, YX; Xu, LW; Ding, QR
联系作者: Ding, QR (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Nutr Sci, CAS Key Lab Nutr & Metab, Shanghai 200031, Peoples R China.
英文联系作者: Ding, QR (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Nutr Sci, CAS Key Lab Nutr & Metab, Shanghai 200031, Peoples R China.
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发表年度: 2017
卷: 23
期: 1
页码: 1-5
摘要: 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 promoterdriven sgRNA expression needs to be reevaluated before it can be used for therapeutic genome editing.
英文摘要: 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 promoterdriven sgRNA expression needs to be reevaluated before it can be used for therapeutic genome editing.
刊物名称: RNA
英文刊物名称: RNA
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学科: Biochemistry & Molecular Biology
英文学科: Biochemistry & Molecular Biology
影响因子: 4.605
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论文类别: Article
英文论文类别: Article
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