Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease
We have developed a CRISPR-based method that uses catalytically active Cas9 and distinct single guide (sgRNA) constructs to knock out and activate different genes in the same cell. These sgRNAs, with 14- to 15-bp target sequences and MS2 binding loops, can activate gene expression using an active St...
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| Format: | Article |
| Language: | en_US |
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Nature Publishing Group
2016
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| Online Access: | http://hdl.handle.net/1721.1/102583 https://orcid.org/0000-0003-2782-2509 https://orcid.org/0000-0001-7915-1685 https://orcid.org/0000-0001-6656-5002 https://orcid.org/0000-0002-7979-3220 |
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| author | Joung, Julia Zhang, Feng Konermann, Silvana Dahlman, James E. Abudayyeh, Omar Osama Gootenberg, Jonathan S. |
| author2 | Harvard University--MIT Division of Health Sciences and Technology |
| author_facet | Harvard University--MIT Division of Health Sciences and Technology Joung, Julia Zhang, Feng Konermann, Silvana Dahlman, James E. Abudayyeh, Omar Osama Gootenberg, Jonathan S. |
| author_sort | Joung, Julia |
| collection | MIT |
| description | We have developed a CRISPR-based method that uses catalytically active Cas9 and distinct single guide (sgRNA) constructs to knock out and activate different genes in the same cell. These sgRNAs, with 14- to 15-bp target sequences and MS2 binding loops, can activate gene expression using an active Streptococcus pyogenes Cas9 nuclease, without inducing double-stranded breaks. We use these 'dead RNAs' to perform orthogonal gene knockout and transcriptional activation in human cells. |
| first_indexed | 2024-09-23T13:21:28Z |
| format | Article |
| id | mit-1721.1/102583 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T13:21:28Z |
| publishDate | 2016 |
| publisher | Nature Publishing Group |
| record_format | dspace |
| spelling | mit-1721.1/1025832022-09-28T13:39:55Z Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease Joung, Julia Zhang, Feng Konermann, Silvana Dahlman, James E. Abudayyeh, Omar Osama Gootenberg, Jonathan S. Harvard University--MIT Division of Health Sciences and Technology Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences McGovern Institute for Brain Research at MIT Abudayyeh, Omar Osama Joung, Julia Gootenberg, Jonathan S. Zhang, Feng Konermann, Silvana We have developed a CRISPR-based method that uses catalytically active Cas9 and distinct single guide (sgRNA) constructs to knock out and activate different genes in the same cell. These sgRNAs, with 14- to 15-bp target sequences and MS2 binding loops, can activate gene expression using an active Streptococcus pyogenes Cas9 nuclease, without inducing double-stranded breaks. We use these 'dead RNAs' to perform orthogonal gene knockout and transcriptional activation in human cells. McGovern Institute for Brain Research at MIT (Friends of McGovern Institute Fellowship) United States. Dept. of Energy (Computational Science Graduate Fellowship) National Institute of Mental Health (U.S.) (1DP1-MH100706) Poitras Foundation Vallee Foundation Simons Foundation Paul G. Allen Foundation New York Stem Cell Foundation 2016-05-22T23:41:33Z 2016-05-22T23:41:33Z 2015-10 2015-08 Article http://purl.org/eprint/type/JournalArticle 1087-0156 1546-1696 http://hdl.handle.net/1721.1/102583 Dahlman, James E, Omar O Abudayyeh, Julia Joung, Jonathan S Gootenberg, Feng Zhang, and Silvana Konermann. “Orthogonal Gene Knockout and Activation with a Catalytically Active Cas9 Nuclease.” Nature Biotechnology 33, no. 11 (October 5, 2015): 1159–61. https://orcid.org/0000-0003-2782-2509 https://orcid.org/0000-0001-7915-1685 https://orcid.org/0000-0001-6656-5002 https://orcid.org/0000-0002-7979-3220 en_US http://dx.doi.org/10.1038/nbt.3390 Nature Biotechnology Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf Nature Publishing Group PMC |
| spellingShingle | Joung, Julia Zhang, Feng Konermann, Silvana Dahlman, James E. Abudayyeh, Omar Osama Gootenberg, Jonathan S. Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease |
| title | Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease |
| title_full | Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease |
| title_fullStr | Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease |
| title_full_unstemmed | Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease |
| title_short | Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease |
| title_sort | orthogonal gene knockout and activation with a catalytically active cas9 nuclease |
| url | http://hdl.handle.net/1721.1/102583 https://orcid.org/0000-0003-2782-2509 https://orcid.org/0000-0001-7915-1685 https://orcid.org/0000-0001-6656-5002 https://orcid.org/0000-0002-7979-3220 |
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