High frequency targeted mutagenesis using engineered endonucleases and DNA-end processing enzymes.
Targeting DNA double-strand breaks is a powerful strategy for gene inactivation applications. Without the use of a repair plasmid, targeted mutagenesis can be achieved through Non-Homologous End joining (NHEJ) pathways. However, many of the DNA breaks produced by engineered nucleases may be subject...
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Public Library of Science (PLoS)
2013
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oai:doaj.org-article:f3cd0e8d8ff24ecf88c47733d9ec49182021-11-18T08:00:09ZHigh frequency targeted mutagenesis using engineered endonucleases and DNA-end processing enzymes.1932-620310.1371/journal.pone.0053217https://doaj.org/article/f3cd0e8d8ff24ecf88c47733d9ec49182013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23359797/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Targeting DNA double-strand breaks is a powerful strategy for gene inactivation applications. Without the use of a repair plasmid, targeted mutagenesis can be achieved through Non-Homologous End joining (NHEJ) pathways. However, many of the DNA breaks produced by engineered nucleases may be subject to precise re-ligation without loss of genetic information and thus are likely to be unproductive. In this study, we combined engineered endonucleases and DNA-end processing enzymes to increase the efficiency of targeted mutagenesis, providing a robust and efficient method to (i) greatly improve targeted mutagenesis frequency up to 30-fold, and; (ii) control the nature of mutagenic events using meganucleases in conjunction with DNA-end processing enzymes in human primary cells.Fabien DelacôteChristophe PerezValérie GuyotMarianne DuhamelChristelle RochonNathalie OllivierRachel MacmasterGeorge H SilvaFrédéric PâquesFayza DaboussiPhilippe DuchateauPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 1, p e53217 (2013) |
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DOAJ |
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DOAJ |
| language |
EN |
| topic |
Medicine R Science Q |
| spellingShingle |
Medicine R Science Q Fabien Delacôte Christophe Perez Valérie Guyot Marianne Duhamel Christelle Rochon Nathalie Ollivier Rachel Macmaster George H Silva Frédéric Pâques Fayza Daboussi Philippe Duchateau High frequency targeted mutagenesis using engineered endonucleases and DNA-end processing enzymes. |
| description |
Targeting DNA double-strand breaks is a powerful strategy for gene inactivation applications. Without the use of a repair plasmid, targeted mutagenesis can be achieved through Non-Homologous End joining (NHEJ) pathways. However, many of the DNA breaks produced by engineered nucleases may be subject to precise re-ligation without loss of genetic information and thus are likely to be unproductive. In this study, we combined engineered endonucleases and DNA-end processing enzymes to increase the efficiency of targeted mutagenesis, providing a robust and efficient method to (i) greatly improve targeted mutagenesis frequency up to 30-fold, and; (ii) control the nature of mutagenic events using meganucleases in conjunction with DNA-end processing enzymes in human primary cells. |
| format |
article |
| author |
Fabien Delacôte Christophe Perez Valérie Guyot Marianne Duhamel Christelle Rochon Nathalie Ollivier Rachel Macmaster George H Silva Frédéric Pâques Fayza Daboussi Philippe Duchateau |
| author_facet |
Fabien Delacôte Christophe Perez Valérie Guyot Marianne Duhamel Christelle Rochon Nathalie Ollivier Rachel Macmaster George H Silva Frédéric Pâques Fayza Daboussi Philippe Duchateau |
| author_sort |
Fabien Delacôte |
| title |
High frequency targeted mutagenesis using engineered endonucleases and DNA-end processing enzymes. |
| title_short |
High frequency targeted mutagenesis using engineered endonucleases and DNA-end processing enzymes. |
| title_full |
High frequency targeted mutagenesis using engineered endonucleases and DNA-end processing enzymes. |
| title_fullStr |
High frequency targeted mutagenesis using engineered endonucleases and DNA-end processing enzymes. |
| title_full_unstemmed |
High frequency targeted mutagenesis using engineered endonucleases and DNA-end processing enzymes. |
| title_sort |
high frequency targeted mutagenesis using engineered endonucleases and dna-end processing enzymes. |
| publisher |
Public Library of Science (PLoS) |
| publishDate |
2013 |
| url |
https://doaj.org/article/f3cd0e8d8ff24ecf88c47733d9ec4918 |
| work_keys_str_mv |
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| _version_ |
1718422704099426304 |