The role and fate of DNA ends for homologous recombination in embryonic stem cells.

Abstract:

:We have analyzed the gene-targeting frequencies and recombination products generated by a series of vectors which target the hprt locus in embryonic stem cells and found the existence of alternative pathways that depend on the location of the double-strand break within the vector. A double-strand break in the targeting homology was found to increase the targeting frequency compared with a double-strand break at the edge of or outside the target homology; this finding agrees with the double-strand break repair model proposed for Saccharomyces cerevisiae. Although a double-strand break in the homology is important for efficient targeting, observations reported here suggest that the terminal ends are not always directly involved in the initial recombination event. Short terminal heterologous sequences which block the homologous ends of the vector may be incorporated into the target locus. A modification of the double-strand break repair model is described to account for this observation.

journal_name

Mol Cell Biol

authors

Hasty P,Rivera-Pérez J,Bradley A

doi

10.1128/mcb.12.6.2464

subject

Has Abstract

pub_date

1992-06-01 00:00:00

pages

2464-74

issue

6

eissn

0270-7306

issn

1098-5549

journal_volume

12

pub_type

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