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Nucleic Acids Research, 1995, Vol. 23, No. 11 2058-2064
© 1995

MOLECULAR BIOLOGY

Gene conversion during vector insertion in embryonic stem cells

Paul Hasty1,*, Jaime Rivera-Pérez1 and Allan Bradley1,2

1 Department of Human and Molecular Genetics USA 2 Howard Hughes Medical Institute, Baylor College of Medicine, One Baylor Plaza Houston, TX 77030, USA

* To whom correspondence should be addressed at present address: The University of Texas, M. D. Anderson Cancer Center, Department of Biochemistry and Molecular Biology, 1515 Holcombe Boulevard, Houston, TX 77030, USA

Received December 22, 1994. Revised April 10, 1995. Accepted April 10, 1995.

Recombination of an insertion vector Into Its chromosomal homologue is a conservative event in that both the chromosomal and the vector sequences are preserved. However, gene conversion may accompany homologous recombination of an Insertion vector. To examine gene conversion in more detail we have determined the targeting frequencies and the structure of the recomblnant alleles generated with a series of vectors which target the hprt gene in embryonic stem cells. We demonstrate that gene conversion of the introduced mutation does not significantly limit homologous recombination and that gene conversion occurs without a sequence specific bias for five different mutations. The frequency of the loss of a vector mutation and the gain of a chromosomal sequence is Inversely proportional to the distance between the vector mutation and the double-strand break. The loss of a chromosomal sequence and the gain of a vector mutation occurs at a low frequency.


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