A system for rapid generation of coat color-tagged knockouts and defined chromosomal rearrangements in mice

Zheng, B., Mills, A. A., Bradley, A. (1999) A system for rapid generation of coat color-tagged knockouts and defined chromosomal rearrangements in mice. Nucleic Acids Research, 27 (11). pp. 2354-2360. ISSN 03051048 (ISSN)

URL: http://www.ncbi.nlm.nih.gov/pubmed/10325425
DOI: 10.1093/nar/27.11.2354

Abstract

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 chromosomal 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.

Item Type: Paper
Uncontrolled Keywords: animal cell article chromosome map chromosome rearrangement embryo cell gene mutation gene targeting genetic engineering genetic marker genome genotype knockout mouse library molecular cloning mouse mutagenesis nonhuman priority journal stem cell Animals Gene Rearrangement Hair Color Intercellular Signaling Peptides and Proteins Mice Mice, Inbred C57BL Mice, Knockout Monophenol Monooxygenase Proteins Time Factors Animalia
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing
bioinformatics > genomics and proteomics
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > chromosome
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > chromosomes, structure and function > chromosome

organism description > animal > mammal > rodent > mouse
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > transgenic animal
CSHL Authors:
Communities: CSHL labs > Mills lab
Depositing User: Matt Covey
Date: 1999
Date Deposited: 11 Mar 2013 20:40
Last Modified: 11 Mar 2013 20:40
PMCID: PMC148802
Related URLs:
URI: http://repository.cshl.edu/id/eprint/27765

Actions (login required)

Administrator's edit/view item Administrator's edit/view item
CSHL HomeAbout CSHLResearchEducationNews & FeaturesCampus & Public EventsCareersGiving