| First Author | Zelensky AN | Year | 2017 |
| Journal | Nat Commun | Volume | 8 |
| Issue | 1 | Pages | 66 |
| PubMed ID | 28687761 | Mgi Jnum | J:249698 |
| Mgi Id | MGI:5920742 | Doi | 10.1038/s41467-017-00124-3 |
| Citation | Zelensky AN, et al. (2017) Inactivation of Pol theta and C-NHEJ eliminates off-target integration of exogenous DNA. Nat Commun 8(1):66 |
| abstractText | Off-target or random integration of exogenous DNA hampers precise genomic engineering and presents a safety risk in clinical gene therapy strategies. Genetic definition of random integration has been lacking for decades. Here, we show that the A-family DNA polymerase theta (Pol theta) promotes random integration, while canonical non-homologous DNA end joining plays a secondary role; cells double deficient for polymerase theta and canonical non-homologous DNA end joining are devoid of any integration events, demonstrating that these two mechanisms define random integration. In contrast, homologous recombination is not reduced in these cells and gene targeting is improved to 100% efficiency. Such complete reversal of integration outcome, from predominately random integration to exclusively gene targeting, provides a rational way forward to improve the efficacy and safety of DNA delivery and gene correction approaches.Random off-target integration events can impair precise gene targeting and poses a safety risk for gene therapy. Here the authors show that repression of polymerase theta and classical non-homologous recombination eliminates random integration. |
