Cas9 gRNA engineering for genome editing, activation and repression

• Main Authors: Chavez, Alejandro (Author), Tuttle, Marcelle (Author), Chari, Raj (Author), Ter-Ovanesyan, Dmitry (Author), Qian, Jason (Author), Pruitt, Benjamin W (Author), Buchthal, Joanna (Author), Church, George (Author), Kiani, Samira (Contributor), Hall, Richard N. (Contributor), Beal, Jacob S (Contributor), Vora, Suhani Deepak (Contributor), Kowal, Emma J. (Contributor), Ebrahimkhani, Mohammad Reza (Contributor), Collins, James J. (Contributor), Weiss, Ron (Contributor)
• Other Authors: MIT Synthetic Biology Center (Contributor), Massachusetts Institute of Technology. Department of Biological Engineering (Contributor)
• Format: Article
• Language: English
• Published: Nature Publishing Group, 2017-04-06T19:57:41Z.
• Subjects: Article
• Online Access: Get fulltext

We demonstrate that by altering the length of Cas9-associated guide RNA(gRNA) we were able to control Cas9 nuclease activity and simultaneously perform genome editing and transcriptional regulation with a single Cas9 protein. We exploited these principles to engineer mammalian synthetic circuits with combined transcriptional regulation and kill functions governed by a single multifunctional Cas9 protein.
National Human Genome Research Institute (U.S.) (P50 HG005550)
United States. Department of Energy (DE-FG02-02ER63445)
Wyss Institute for Biologically Inspired Engineering
United States. Army Research Office (DARPA W911NF-11-2-0054)
National Science Foundation (U.S.)
United States. National Institutes of Health (5R01CA155320-04)
United States. National Institutes of Health (P50 GM098792)
National Cancer Institute (U.S.) (5T32CA009216-34)
Massachusetts Institute of Technology. Department of Biological Engineering
Harvard Medical School. Department of Genetics
Defense Threat Reduction Agency (DTRA) (HDTRA1-14-1-0006)