Cell-Type-Specific CRISPR Activation with MicroRNA-Responsive AcrllA4 Switch

被引:40
作者
Hirosawa, Moe [1 ,2 ]
Fujita, Yoshihiko [1 ]
Saito, Hirohide [1 ]
机构
[1] Kyoto Univ, Ctr iPS Cell Res & Applicat CiRA, Dept Life Sci Frontiers, Kyoto 6068501, Japan
[2] Kyoto Univ, Grad Sch Med, Kyoto 6068501, Japan
来源
ACS SYNTHETIC BIOLOGY | 2019年 / 8卷 / 07期
基金
日本学术振兴会;
关键词
CRISPR-Cas9; anti-CRISPR; synthetic biology; microRNA; RNA switch; GENOME; INHIBITION; GENE;
D O I
10.1021/acssynbio.9b00073
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Anti-CRISPR proteins have the potential to regulate CRISPR-Cas systems in a cell-type-specific manner. To selectively edit the genome in target cells, we controlled the expression of AcrllA4, a Streptococcus pyogenes Cas9 inhibitor, based on endogenous microRNA (miRNA) activity. We designed a miRNA-responsive AcrllA4 switch, which is a synthetic mRNA that contains a completely complementary sequence to an arbitrary miRNA at the 5'-UTR region and encodes AcrllA4. Together with the Cas9- or dCas9-VPR-guide RNA complex, this switch functions as a cell-specific Cas9 or dCas9-VPR activator that induces gene knockout or activation depending on the target miRNA. By sensing intracellular miRNAs, the conditional CRISPR-Cas9 ON system that we report could provide a powerful tool for future therapeutic applications and genome engineering.
引用
收藏
页码:1575 / 1582
页数:15
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