Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 115, Issue 29, Pages E6722-E6730Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1802448115
Keywords
synthetic biology; optogenetics; far-red light; CRISPR-dCas9; regenerative medicine
Categories
Funding
- National Natural Science Foundation of China (NSFC) [31522017]
- National Key Research and Development Program of China, Stem Cell and Translational Research Grant [2016YFA0100300]
- NSFC [31470834, 31670869]
- Science and Technology Commission of Shanghai Municipality [18JC1411000]
- Thousand Youth Talents Plan of China
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The ability to control the activity of CRISPR-dCas9 with precise spatiotemporal resolution will enable tight genome regulation of user-defined endogenous genes for studying the dynamics of transcriptional regulation. Optogenetic devices with minimal photo-toxicity and the capacity for deep tissue penetration are extremely useful for precise spatiotemporal control of cellular behavior and for future clinic translational research. Therefore, capitalizing on synthetic biology and optogenetic design principles, we engineered a far-red light (FRL)-activated CRISPR-dCas9 effector (FACE) device that induces transcription of exogenous or endogenous genes in the presence of FRL stimulation. This versatile system provides a robust and convenient method for precise spatiotemporal control of endogenous gene expression and also has been demonstrated to mediate targeted epigenetic modulation, which can be utilized to efficiently promote differentiation of induced pluripotent stem cells into functional neurons by up-regulating a single neural transcription factor, NEUROG2. This FACE system might facilitate genetic/epigenetic reprogramming in basic biological research and regenerative medicine for future biomedical applications.
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