Journal
ACS SYNTHETIC BIOLOGY
Volume 6, Issue 4, Pages 686-693Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.6b00313
Keywords
CRISPR/Cas9; synthetic transcription factor; chemically induced dimerization; genetic regulation; orthogonality
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Funding
- Carl R. Woese Institute for Genomic Biology at the University of Illinois at Urbana-Champaign
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The concerted action of multiple genes in a time-dependent manner controls complex cellular phenotypes, yet the temporal regulation of gene expressions is restricted on a single-gene level, which limits our ability to control higher-order gene networks and understand the consequences of multiplex genetic perturbations. Here we developed a system for temporal regulation of multiple genes. This system combines the simplicity of CRISPR/Cas9 activators for orthogonal targeting of multiple genes and the orthogonality of chemically induced dimerizing (CID) proteins for temporal control of CRISPR/Cas9 activator function. In human cells, these transcription activators exerted simultaneous activation of multiple genes and orthogonal regulation of different genes in a ligand-dependent manner with minimal background. We envision that our system will enable the perturbation of higher-order gene networks with high temporal resolution and accelerate our understanding of gene gene interactions in a complex biological setting.
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