Rewiring Calcium Signaling for Precise Transcriptional Reprogramming

ACS Synthetic Biology
Nhung Thi NguyenYubin Zhou

Abstract

Tools capable of modulating gene expression in living organisms are very useful for interrogating the gene regulatory network and controlling biological processes. The catalytically inactive CRISPR/Cas9 (dCas9), when fused with repressive or activating effectors, functions as a versatile platform to reprogram gene transcription at targeted genomic loci. However, without temporal control, the application of these reprogramming tools will likely cause off-target effects and lack strict reversibility. To overcome this limitation, we report herein the development of a chemical or light-inducible transcriptional reprogramming device that combines photoswitchable genetically encoded calcium actuators with dCas9 to control gene expression. By fusing an engineered Ca2+-responsive NFAT fragment with dCas9 and transcriptional coactivators, we harness the power of light to achieve photoinducible transcriptional reprogramming in mammalian cells. This synthetic system (designated CaRROT) can also be used to document calcium-dependent activity in mammals after exposure to ligands or chemicals that would elicit calcium response inside cells.

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Citations

Jul 25, 2018·Critical Reviews in Biochemistry and Molecular Biology·Robert M Hughes
Jan 12, 2020·Nature Communications·Sungsoo KimWon Do Heo
Jul 14, 2020·Biochemical Society Transactions·Denis HartmannMichael J Booth
Feb 1, 2020·Nature Communications·Krzysztof KrawczykMartin Fussenegger
Feb 2, 2021·ACS Biomaterials Science & Engineering·Ross C Bretherton, Cole A DeForest
Nov 24, 2020·Current Opinion in Physiology·Marc FahrnerChristoph Romanin
Jan 13, 2021·Nature Communications·Lian HeYubin Zhou
Dec 9, 2020·The FEBS Journal·Anže VerbičRoman Jerala
Oct 12, 2018·Cell Reports·Mayumi YamadaItaru Imayoshi
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Jul 10, 2020·ACS Synthetic Biology·Maja MeškoMojca Benčina
Apr 12, 2020·Journal of Molecular Biology·Yi-Tsang LeeYubin Zhou

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