Hunt, Victoria M.Chen, Wilfred2022-05-272022-05-272022-04-26Hunt, Victoria M., and Wilfred Chen. 2022. “A MicroRNA-Gated ThgRNA Platform for Multiplexed Activation of Gene Expression in Mammalian Cells.” Chemical Communications 58 (42): 6215–18. https://doi.org/10.1039/D2CC01478E.1364-548Xhttps://udspace.udel.edu/handle/19716/30910This article was originally published in Chemical Communications. The version of record is available at: https://doi.org/10.1039/D2CC01478E. This article will be embargoed until 04/26/2023.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.en-USArticle