Mechanisms involved in targeted gene replacement in mammalian cells.

摘要: The "ends-out" or omega (Omega)-form gene replacement vector is used routinely to perform targeted genome modification in a variety of species and has the potential be an effective vehicle for therapy. However, mammalian cells, frequency this reaction low mechanism unknown. Understanding molecular features associated with important may lead increase efficiency process. In study, we investigated cells using powerful assay system that permits efficient recovery product(s) individual recombination events at haploid, chromosomal mu-delta locus murine hybridoma cell line. results showed (i) heteroduplex DNA (hDNA) formed during replacement; (ii) mismatches hDNA are usually efficiently repaired before replication division; (iii) occurs fidelity; (iv) presence multiple markers one homologous flanking arm did not affect (v) comparison genomic fragment bearing contiguous homology target, targeting was only slightly inhibited by internal heterology (pSV2neo sequences) vector.