Cas9 gRNA engineering for genome editing, activation and repression.

Autor: Kiani S; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.; Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Chavez A; Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA.; Department of Pathology, Massachusetts General Hospital, Boston, Massachusetts, USA.; Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA., Tuttle M; Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA., Hall RN; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.; Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Chari R; Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA., Ter-Ovanesyan D; Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA.; Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA., Qian J; Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA.; Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA., Pruitt BW; Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA., Beal J; Raytheon BBN Technologies, Cambridge, Massachusetts, USA., Vora S; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.; Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA.; Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA., Buchthal J; Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA., Kowal EJ; Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA., Ebrahimkhani MR; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.; Center for Emergent Behaviors of Integrated Cellular Systems (EBICS), Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Collins JJ; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.; Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.; Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA.; Institute for Medical Engineering &Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.; Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.; Harvard-MIT Program in Health Sciences and Technology, Cambridge, Massachusetts, USA., Weiss R; Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.; Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.; Center for Emergent Behaviors of Integrated Cellular Systems (EBICS), Massachusetts Institute of Technology, Cambridge, Massachusetts, USA., Church G; Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, Massachusetts, USA.; Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
Jazyk: angličtina
Zdroj: Nature methods [Nat Methods] 2015 Nov; Vol. 12 (11), pp. 1051-4. Date of Electronic Publication: 2015 Sep 07.
DOI: 10.1038/nmeth.3580
Abstrakt: 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.
Databáze: MEDLINE