Activatable CRISPR Transcriptional Circuits Lighted up by Fluorescent RNA for mRNA Sensing and Silencing.
Activatable CRISPR Transcriptional Circuits Lighted up by Fluorescent RNA for mRNA Sensing and Silencing.
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DOI：
10.1002/anie.202004751
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发表时间：
2020-07
影响因子：
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通讯作者：
Zhan‐Ming Ying;Fenglin Wang;X. Chu;R. Yu;Jian-hui Jiang
中科院分区：
文献类型：
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作者：
Zhan‐Ming Ying;Fenglin Wang;X. Chu;R. Yu;Jian-hui Jiang
用细胞特异性信息精确激活的CRISPR-dCas 9系统提高了开发智能传感器或治疗策略的能力。我们报告了可激活的dCas 9转录电路的首次开发，该转录电路能够使用杂交介导的gRNA激活结构转换来感测和沉默活细胞中的mRNA。gRNA被设计为具有被发夹结构阻断的间隔序列，并且mRNA杂交诱导gRNA结构转换并激活报告RNA的转录。使用点亮RNA报告基因开发的mRNA传感器显示出对药物处理下的细胞和不同细胞系中的生存素mRNA的高灵敏度和快速响应成像。此外，通过在报告RNA中掺入小发夹RNA来工程化反馈回路。它展示了一个智能的策略，动态传感和沉默生存素诱导肿瘤细胞凋亡。该电路说明了一个广泛适用的平台，以开发细胞特异性传感和治疗策略。
CRISPR-dCas9 systems precisely activated with cell-specific information improve abilities to develop smart sensors or therapeutic strategies. We report the first development of an activatable dCas9 transcriptional circuit that enables sensing and silencing of mRNA in living cells using hybridization-mediated structure switching for gRNA activation. The gRNA is designed with the spacer sequence blocked by a hairpin structure, and mRNA hybridization induces gRNA structure switching and activates transcription of reporter RNA. A mRNA sensor developed using light-up RNA reporter shows high sensitivity and fast-response imaging of survivin mRNA in cells under drug treatments and different cell lines. Furthermore, a feedback circuit is engineered by incorporating a small hairpin RNA in reporter RNA. It demonstrates a smart strategy for dynamic sensing and silencing of survivin with induced tumor cell apoptosis. This circuit illustrates a broadly applicable platform to develop cell-specific sensing and therapeutic strategies.