Cas9 gRNA engineering for genome editing, activation and repression
| Autor: | George M. Church, Richard N. Hall, Benjamin W. Pruitt, Suhani Vora, Mohammad R. Ebrahimkhani, Dmitry Ter-Ovanesyan, Jacob Beal, Alejandro Chavez, Samira Kiani, Ron Weiss, Emma J. K. Kowal, James J. Collins, Marcelle Tuttle, Joanna Buchthal, Jason Qian, Raj Chari |
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| Přispěvatelé: | MIT Synthetic Biology Center, Massachusetts Institute of Technology. Department of Biological Engineering, Kiani, Samira, Hall, Richard N., Beal, Jacob S, Vora, Suhani Deepak, Kowal, Emma J., Ebrahimkhani, Mohammad Reza, Collins, James J., Weiss, Ron |
| Rok vydání: | 2015 |
| Předmět: |
Transcription
Genetic CRISPR-Associated Proteins Genetic Vectors Computational biology Biology Biochemistry Genome Article Genome editing Genes Reporter Transcriptional regulation Humans Guide RNA Molecular Biology Gene Fluorescent Dyes Genetics Regulation of gene expression Binding Sites Cas9 Cell Biology Flow Cytometry HEK293 Cells Microscopy Fluorescence Mutagenesis RNA editing Mutation RNA Editing CRISPR-Cas Systems Genetic Engineering Gene Deletion RNA Guide Kinetoplastida Biotechnology |
| Zdroj: | PMC |
| ISSN: | 1548-7105 1548-7091 |
| Popis: | 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) |
| Databáze: | OpenAIRE |
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