A microRNA-gated thgRNA platform for multiplexed activation of gene expression in mammalian cells. Issue 42 (4th May 2022)

• Record Type: Journal Article
• Authors: Hunt, Victoria M.; Chen, Wilfred
• Is Part Of: Chemical communications. Volume 58:Issue 42(2022)
• Page Start: 6215
• Page End: 6218
• Publication Date: 2022-05-04
• Subjects: Chemistry -- Periodicals; 540
• DOI: 10.1039/d2cc01478e
• Languages: English
• ISSNs: 1359-7345

Abstract

Abstract : Using a newly created AND-gate toehold-gated guide RNA (thgRNA), presence of specific miRNA and mRNA allows conditional gene activation and protein degradation. Abstract : To effectively reprogram cellular regulatory networks towards desired phenotypes, it is critical to have the ability to provide precise gene regulation in a spatiotemporal manner. We have previously engineered toehold-gated guide RNA (thgRNA) to enable conditional activation of dCas9-mediated transcriptional upregulation in mammalian cells using synthetic RNA triggers. Here, we demonstrate that microRNA (miR)-gated thgRNAs can be transcribed by type II RNA polymerase to allow multiplexed transcriptional activation using both mRNA and miR. Activation is achieved only by proper miR-mediated processing of the flanking 5′ cap and 3′ poly A tail and hairpin unblocking by mRNA via strand displacement. This new AND-gate design is exploited to elicit conditional protein degradation based on induced expression of a specific ubiquibody. This new strategy may find many new applications in an RNA-responsive manner.