Incorporation of Large Heterologies Into Heteroduplex DNA During Double-Strand-Break Repair in Mouse Cells

Incorporation of Large Heterologies Into Heteroduplex DNA During Double-Strand-Break Repair in Mouse Cells

Steven J. Raynard^a and Mark D. Baker^a,b
^a Department of Molecular Biology and Genetics, College of Biological Science
^b Department of Pathobiology, ppOntario Veterinary College, University of Guelph, Guelph, Ontario N1G 2W1, Canada

Corresponding author: Mark D. Baker, Ontario Veterinary College, University of Guelph, Guelph, ON N1G 2W1, Canada., mdbaker{at}uoguelph.ca (E-mail)

Communicating editor: L. S. SYMINGTON

In this study, the formation and repair of large (>1 kb) insertion/deletion(I/D) heterologies during double-strand-break repair (DSBR)was investigated using a gene-targeting assay that permits efficientrecovery of sequence insertion events at the haploid chromosomalimmunoglobulin (Ig) µ-locus in mouse hybridoma cells.The results revealed that (i) large I/D heterologies were generatedon one or both sides of the DSB and, in some cases, formed symmetricallyin both homology regions; (ii) large I/D heterologies did notnegatively affect the gene targeting frequency; and (iii) priorto DNA replication, the large I/D heterologies were rectified.

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