Nucleic Acids Research, Vol 27, Issue 11 2354-2360, Copyright © 1999 by Oxford University Press
B Zheng, AA Mills and A Bradley
Gene targeting in mouse embryonic stem (ES) cells can be used to generate
single gene mutations or defined multi-megabase chromosomal rearrangements
when applied with the Cre- loxP recombination system. While single
knockouts are essential for uncovering functions of cloned genes,
chromosomal rearrangements are great genetic tools for mapping, mutagenesis
screens and functional genomics. The conventional approach to generate mice
with targeted alterations of the genome requires extensive molecular
cloning to build targeting vectors and DNA-based genotyping for stock
maintenance. Here we describe the design and construction of a two-library
system to facilitate high throughput gene targeting and chromo-somal
engineering. The unique feature of these libraries is that once a clone is
isolated, it is essentially ready to be used for insertional targeting in
ES cells. The two libraries each bear a complementary set of genetic
markers tailored so that the vector can be used for Cre- loxP -based
chromosome engineering as well as single knockouts. By incorporating mouse
coat color markers into the vectors, we illustrate a widely applicable
method for stock maintenance of ES cell-derived mice with single gene
knockouts or more extensive chromosomal rearrangements.
ARTICLES
A system for rapid generation of coat color-tagged knockouts and defined chromosomal rearrangements in mice
Department of Molecular and Human Genetics and Howard Hughes Medical Institute, Baylor College of Medicine,1 Baylor Plaza, Houston, TX 77030, USA.
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