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PMID: 1588950 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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

Molecular and cellular biology ·Vol. 12 ·No. 6 ·1992-06-00 ·Pages 2464-74

Hasty P, Rivera-Pérez J, Bradley A

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.

MeSH Terms
Animals Base Sequence Cells, Cultured DNA/chemistry DNA, Circular/chemistry DNA, Superhelical/chemistry Genetic Vectors Hypoxanthine Phosphoribosyltransferase/genetics In Vitro Techniques Mice Molecular Sequence Data Oligodeoxyribonucleotides/chemistry Polymerase Chain Reaction Recombination, Genetic Sequence Homology, Nucleic Acid Transfection
Chemicals
DNA, Circular DNA, Superhelical Oligodeoxyribonucleotides DNA Hypoxanthine Phosphoribosyltransferase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Hasty P
Institute for Molecular Genetics, Baylor College of Medicine, Houston, Texas 77030.
Rivera-Pérez J
Bradley A
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.12.6.2464-2474.1992
Published
1992-06-00
Pages
2464-74
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC364439
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.12.6.2464-2474.1992 PMC Full Text View journal details
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