Cell-Type-Specific CRISPR Activation with MicroRNA-Responsive AcrllA4 Switch
Cell-Type-Specific CRISPR Activation with MicroRNA-Responsive AcrllA4 Switch
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DOI：
10.1021/acssynbio.9b00073
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发表时间：
2019-07-01
影响因子：
4.7
通讯作者：
Saito, Hirohide
中科院分区：
文献类型：
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作者：
Hirosawa, Moe;Fujita, Yoshihiko;Saito, Hirohide
抗CRISPR蛋白具有以细胞类型特异性方式调节CRISPR-Cas系统的潜力。为了选择性地编辑靶细胞中的基因组，我们基于内源性microRNA（miRNA）活性控制AcrllA 4（一种化脓性链球菌Cas9抑制剂）的表达。我们设计了一个miRNA响应的AcrllA 4开关，它是一种合成的mRNA，在5 '-UTR区域含有与任意miRNA完全互补的序列，并编码AcrllA 4。与Cas9-或dCas 9-VPR-指导RNA复合物一起，该开关作为细胞特异性Cas9或dCas 9-VPR激活剂起作用，其根据靶miRNA诱导基因敲除或激活。通过检测细胞内的miRNA，我们报告的条件CRISPR-Cas9 ON系统可以为未来的治疗应用和基因组工程提供强大的工具。
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.