Impact of non-homologous end-joining deficiency on random and targeted DNA integration: implications for gene targeting.

ABSTRACT: In higher animal cells, the principal limitation of gene-targeting technology is the extremely low efficiency of targeted integration, which occurs three to four orders of magnitude less frequently than random integration. Assuming that random integration mechanistically involves non-homologous end-joining (NHEJ), inactivation of this pathway should reduce random integration and may enhance gene targeting. To test this possibility, we examined the frequencies of random and targeted integration in NHEJ-deficient chicken DT40 and human Nalm-6 cell lines. As expected, loss of NHEJ resulted in drastically reduced random integration in DT40 cells. Unexpectedly, however, this was not the case for Nalm-6 cells, indicating that NHEJ is not the sole mechanism of random integration. Nevertheless, we

SUBMITTER: Iiizumi S 

PROVIDER: S-EPMC2577324 | biostudies-literature | 2008 Nov

REPOSITORIES: biostudies-literature