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

Paul Hasty, Jaime Rivera-Pérez, Allan Bradley

Research output: Contribution to journalArticlepeer-review

47 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)2464-2474
Number of pages11
JournalMolecular and cellular biology
Volume12
Issue number6
DOIs
StatePublished - Jun 1992

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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