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ACS Synth Biol. 2018 Mar 16;7(3):814-821. doi: 10.1021/acssynbio.7b00467. Epub 2018 Mar 6.

Rewiring Calcium Signaling for Precise Transcriptional Reprogramming.

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.

KEYWORDS:

CRISPR/Cas9; NFAT; calcium signaling; genome editing; optogenetics; transcriptional reprogramming

PMID:
29489336
DOI:
10.1021/acssynbio.7b00467
[Indexed for MEDLINE]

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