RNA Strand Displacement Responsive CRISPR/Cas9 System for mRNA Sensing

Analytical Chemistry
Yue LiJinghong Li

Abstract

CRISPR/Cas9 has already become a powerful tool for genomic manipulation, and further engineering of the system allows it to be precisely regulated in response to external signals, thus, broadening its application possibilities, such as biosensing or bioimaging. However, most stimuli-responsive CRISPR systems are built based on elaborately designed and engineered inducible Cas9 proteins, and external stimuli are still mostly limited as small molecules and light. To construct more precise and easy-to-build responsive CRISPR systems and broaden their responsive species, we seek to engineer conditional guide RNA, rather than Cas9 protein, to mediate conditional CRISPR corresponding to logic operation. Here, we construct mRNA-sensing CRISPR by gRNA reconfiguration and toehold mediated strand displacement, in which each target site could be independently controlled. We show that switches can be embedded into the gRNA and used as RNA sensors, capable of detecting multiple mRNA inputs orthogonally and providing CRISPR/Cas9 response outputs. NOR and NAND logical gates are also constructed, demonstrating its orthogonality and programmability. This strategy promises potential uses in constructing genetic circuits to detect endogenous mRNA...Continue Reading

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Citations

Jan 15, 2020·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Zhifeng ZhaoJinghong Li
Feb 29, 2020·Chemical Society Reviews·Kaixiang ZhangJinghong Li
Mar 17, 2020·Biotechnology and Bioengineering·Chu-Shu ZhuLv-Yun Zhu
Jul 4, 2019·Current Gene Therapy·Cia-Hin Lau, Chung Tin
Nov 16, 2019·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Yiting WangJianhua Wang
Oct 24, 2020·The CRISPR Journal·Roberto GaliziAlfonso Jaramillo
Nov 18, 2020·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Yijing JiaJin Ouyang
Feb 2, 2020·Bioelectrochemistry·Zahra HatamiMojtaba Shamsipur
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Aug 14, 2019·Analytical Chemistry·Mingshu XiaoHao Pei
Sep 19, 2019·ACS Applied Materials & Interfaces·Piaopiao ChenJia Geng
Jun 11, 2020·Analytical Chemistry·Peng GaoBo Tang
Jul 26, 2019·Analytical Chemistry·Jianyu HuYi Lv
Oct 5, 2021·Chemical Society Reviews·Lanlan ChenHuanghao Yang

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