A light-inducible CRISPR-Cas9 system for control of endogenous gene activation

Nature Chemical Biology volume 11, pages198–200(2015)Cite this article

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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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Figure 1: Light-inducible, RNA-guided activation of endogenous human genes by the LACE system.
Figure 2: Dynamic spatial and temporal transcriptional control using the LACE system.

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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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Affiliations

  1. Department of Biomedical Engineering, Duke University, Durham, North Carolina, USA

    Lauren R Polstein & Charles A Gersbach

  2. Center for Genomic and Computational Biology, Duke University, Durham, North Carolina, USA

    Charles A Gersbach

  3. Department of Orthopaedic Surgery, Duke University Medical Center, Durham, North Carolina, USA

    Charles A Gersbach

Authors
  1. Lauren R Polstein
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  2. Charles A Gersbach
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Contributions

L.R.P. and C.A.G. designed experiments, analyzed the data and wrote the manuscript. L.R.P. performed experiments.

Corresponding author

Correspondence to Charles A Gersbach.

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The authors declare no competing financial interests.

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Supplementary Results and Supplementary Figures 1–6. (PDF 1142 kb)

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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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