1. Programmable Transcriptional Modulation with a Structured RNA-Mediated CRISPR-dCas9 Complex
- Author
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Miao He, Xiang Zhou, Zhigang Li, Xue Yin, Wenjie Han, Junxiang Zhou, Xiaoyun Sun, Xiaoyu Liu, Dongbao Yao, and Haojun Liang
- Subjects
Transcriptional Activation ,Colloid and Surface Chemistry ,RNA ,Clustered Regularly Interspaced Short Palindromic Repeats ,General Chemistry ,CRISPR-Cas Systems ,Biochemistry ,Catalysis - Abstract
Multi-module dCas9 engineering systems have been developed for controllable transcriptional manipulation such as chemical- or light-induced systems. However, there is still a need for a separate module that can be used for internal control over the CRISPR-dCas9 system. Here, we describe a multi-module CRISPR-dCas9 system in which a separate structured RNA was applied as a programmable component that could control dCas9-based gene regulation and achieved a higher activation efficiency than dCas9-VPR that is traditionally used. By introducing a microRNA sensor, we generated a dCas9-based transcriptional regulation platform that responded to endogenous microRNAs and allowed controllable activation of endogenous genes. Moreover, we applied the platform to selectively identify HCT116 cells in a cell mixture. This work provides a flexible platform for efficient and controllable gene regulation based on CRISPR-dCas9.
- Published
- 2022