Abstract
Optogenetic systems enable precise spatial and temporal control of cell behavior. We engineered a light-activated CRISPR-Cas9 effector (LACE) system that induces transcription of endogenous genes in the presence of blue light. This was accomplished by fusing the light-inducible heterodimerizing proteins CRY2 and CIB1 to a transactivation domain and the catalytically inactive dCas9, respectively. The versatile LACE system can be easily directed to new DNA sequences for the dynamic regulation of endogenous genes.
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Acknowledgements
C. Tucker (University of Colorado–Denver) provided CRY2 and CIBN plasmids. P. Vosburgh designed and fabricated the three-dimensional photomasks. This work was supported by a US National Institutes of Health (NIH) Director–s New Innovator Award (DP2OD008586), US National Science Foundation Faculty Early Career Development Award (CBET-1151035), NIH R01DA036865, NIH R03AR061042, NIH P30AR066527, and an American Heart Association Scientist Development Grant (10SDG3060033 to C.A.G.). L.R.P. was supported by an American Heart Association Predoctoral Fellowship and NIH Biotechnology Training Grant (T32GM008555).
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L.R.P. and C.A.G. designed experiments, analyzed the data and wrote the manuscript. L.R.P. performed experiments.
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Polstein, L., Gersbach, C. A light-inducible CRISPR-Cas9 system for control of endogenous gene activation. Nat Chem Biol 11, 198–200 (2015). https://doi.org/10.1038/nchembio.1753
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DOI: https://doi.org/10.1038/nchembio.1753
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