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PMID: 2120706 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Introduction of a lacZ reporter gene into the mouse int-2 locus by homologous recombination.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 87 ·No. 19 ·1990-10-00 ·Pages 7688-92

Mansour SL, Thomas KR, Deng CX, Capecchi MR

Abstract

We demonstrate that the frequency of gene targeting is unaffected by the length of nonhomologous DNA transferred to a target chromosomal sequence. A result of this finding is that a much wider spectrum of designed genomic alterations is now feasible. As a first application, we inserted a 5.4-kilobase cassette of nonhomologous DNA into the int-2 locus in mouse embryo-derived stem cells by gene targeting. The inserted DNA contained a lacZ gene positioned to create an in-frame fusion with the int-2 protein-coding region. Upon differentiation of these cells to embryoid bodies, the int-2-lacZ fusion faithfully reproduced the expression pattern of int-2 RNA. This ability to target reporter genes, such as lacZ, to specific mouse loci, combined with the ability to move the tagged gene into different mutant backgrounds, may provide an ideal approach for analyzing interactions among genes that participate in a developmental network.

Related Genes
int-2 lacZ
MeSH Terms
Animals Blotting, Southern Cell Differentiation Cell Line Cell Survival Electric Stimulation Embryo, Mammalian Fluorescent Antibody Technique Genetic Vectors Hypoxanthine Phosphoribosyltransferase/genetics Introns Mice Recombination, Genetic Restriction Mapping Transfection beta-Galactosidase/genetics
Chemicals
Hypoxanthine Phosphoribosyltransferase beta-Galactosidase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Mansour S L
Howard Hughes Medical Institute, Department of Biology, University of Utah, Salt Lake City 84112.
Thomas K R
Deng C X
Capecchi M R
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.87.19.7688
Published
1990-10-00
Pages
7688-92
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC54813
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.87.19.7688 PMC Full Text View journal details
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