Location of crossovers during gene targeting with insertion and replacement vectors.

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Publication Type Journal Article
School or College College of Science; School of Medicine
Department Oncological Sciences; Biology; Human Genetics
Program Institute of Human Genetics; Howard Hughes Medical Institute (HHMI)
Creator Thomas, Kirk R.; Capecchi, Mario R.
Other Author Deng, Chuxia
Title Location of crossovers during gene targeting with insertion and replacement vectors.
Date 1993-04
Description Gene targeting was used to introduce nonselectable genetic changes into chromosomal loci in mouse embryo-derived stem cells. The nonselectable markers were linked to a selectable marker in both insertion- and replacement-type vectors, and the transfer of the two elements to the Hprt locus was assayed. When insertion vectors were used as substrates, the frequency of transfer was highly dependent upon the distance between the nonselectable marker and the double-strand break in the vector. A marker located close to the vector ends was frequently lost, suggesting that a double-strand gap repair activity is involved in vector integration. When replacement vectors were used, cotransfer of a selectable marker and a nonselectable marker 3 kb apart was over 50%, suggesting that recombination between vector and target often occurs near the ends of the vector. To illustrate the use of replacement vectors to transfer specific mutations to the genome, we describe targeting of the delta F508 mutation to the CFTR gene in mouse embryo-derived stem cells.
Type Text
Publisher American Society for Microbiology
Volume 13
Issue 4
First Page 2134
Last Page 2140
Subject Genetic Vectors; Molecular Sequence Data; Restriction Mapping
Subject MESH Mutagenesis, Insertional; Recombination, Genetic; Transfection
Language eng
Bibliographic Citation Mol Cell Biol. 1993 Apr;13(4):2134-40: Deng C, Thomas KR, Capecchi MR. Location of crossovers during gene targeting with insertion and replacement vectors. Retrieved on September 18.2006 from http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=8455602.
Rights Management Copyright © 1993, American Society for Microbiology. All Rights Reserved..
Format Medium application/pdf
Identifier ir-main,432
ARK ark:/87278/s6931bhk
Setname ir_uspace
ID 704415
Reference URL https://collections.lib.utah.edu/ark:/87278/s6931bhk
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