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Highly Efficient Cpf1-Mediated Gene Targeting in Mice Following High Concentration Pronuclear Injection
Cpf1 has emerged as an alternative to the Cas9 RNA-guided nuclease. Here we show that gene targeting rates in mice using Cpf1 can meet, or even surpass, Cas9 targeting rates (approaching 100% targeting), but require higher concentrations of mRNA and guide. We also demonstrate that coinjecting two gu...
| Autores principales: | , , , , , , , , , , |
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| Formato: | Online Artículo Texto |
| Lenguaje: | English |
| Publicado: |
Genetics Society of America
2016
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| Materias: | |
| Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5295614/ https://www.ncbi.nlm.nih.gov/pubmed/28040780 http://dx.doi.org/10.1534/g3.116.038091 |
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| author | Watkins-Chow, Dawn E. Varshney, Gaurav K. Garrett, Lisa J. Chen, Zelin Jimenez, Erin A. Rivas, Cecilia Bishop, Kevin S. Sood, Raman Harper, Ursula L. Pavan, William J. Burgess, Shawn M. |
| author_facet | Watkins-Chow, Dawn E. Varshney, Gaurav K. Garrett, Lisa J. Chen, Zelin Jimenez, Erin A. Rivas, Cecilia Bishop, Kevin S. Sood, Raman Harper, Ursula L. Pavan, William J. Burgess, Shawn M. |
| author_sort | Watkins-Chow, Dawn E. |
| collection | PubMed |
| description | Cpf1 has emerged as an alternative to the Cas9 RNA-guided nuclease. Here we show that gene targeting rates in mice using Cpf1 can meet, or even surpass, Cas9 targeting rates (approaching 100% targeting), but require higher concentrations of mRNA and guide. We also demonstrate that coinjecting two guides with close targeting sites can result in synergistic genomic cutting, even if one of the guides has minimal cutting activity. |
| format | Online Article Text |
| id | pubmed-5295614 |
| institution | National Center for Biotechnology Information |
| language | English |
| publishDate | 2016 |
| publisher | Genetics Society of America |
| record_format | MEDLINE/PubMed |
| spelling | pubmed-52956142017-02-09 Highly Efficient Cpf1-Mediated Gene Targeting in Mice Following High Concentration Pronuclear Injection Watkins-Chow, Dawn E. Varshney, Gaurav K. Garrett, Lisa J. Chen, Zelin Jimenez, Erin A. Rivas, Cecilia Bishop, Kevin S. Sood, Raman Harper, Ursula L. Pavan, William J. Burgess, Shawn M. G3 (Bethesda) Investigations Cpf1 has emerged as an alternative to the Cas9 RNA-guided nuclease. Here we show that gene targeting rates in mice using Cpf1 can meet, or even surpass, Cas9 targeting rates (approaching 100% targeting), but require higher concentrations of mRNA and guide. We also demonstrate that coinjecting two guides with close targeting sites can result in synergistic genomic cutting, even if one of the guides has minimal cutting activity. Genetics Society of America 2016-12-30 /pmc/articles/PMC5295614/ /pubmed/28040780 http://dx.doi.org/10.1534/g3.116.038091 Text en Copyright © 2017 Watkins-Chow et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
| spellingShingle | Investigations Watkins-Chow, Dawn E. Varshney, Gaurav K. Garrett, Lisa J. Chen, Zelin Jimenez, Erin A. Rivas, Cecilia Bishop, Kevin S. Sood, Raman Harper, Ursula L. Pavan, William J. Burgess, Shawn M. Highly Efficient Cpf1-Mediated Gene Targeting in Mice Following High Concentration Pronuclear Injection |
| title | Highly Efficient Cpf1-Mediated Gene Targeting in Mice Following High Concentration Pronuclear Injection |
| title_full | Highly Efficient Cpf1-Mediated Gene Targeting in Mice Following High Concentration Pronuclear Injection |
| title_fullStr | Highly Efficient Cpf1-Mediated Gene Targeting in Mice Following High Concentration Pronuclear Injection |
| title_full_unstemmed | Highly Efficient Cpf1-Mediated Gene Targeting in Mice Following High Concentration Pronuclear Injection |
| title_short | Highly Efficient Cpf1-Mediated Gene Targeting in Mice Following High Concentration Pronuclear Injection |
| title_sort | highly efficient cpf1-mediated gene targeting in mice following high concentration pronuclear injection |
| topic | Investigations |
| url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5295614/ https://www.ncbi.nlm.nih.gov/pubmed/28040780 http://dx.doi.org/10.1534/g3.116.038091 |
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