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PMID: 2821545 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Accurate modification of a chromosomal plasmid by homologous recombination in human cells.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 84 ·No. 19 ·1987-10-00 ·Pages 6820-4

Song KY, Schwartz F, Maeda N, Smithies O, Kucherlapati R

Abstract

We have examined the consequences of modifying mammalian cellular DNA sequences by homologous recombination. A plasmid carrying a 248-base-pair deletion in the neomycin phosphotransferase (neo) gene was introduced into hamster and human cells. The integrated, defective neo gene was used as a target for modification by a second round of transfection with a plasmid carrying a different (283-base-pair) deletion in the neo gene. Recombinants resulting in an intact neo gene were selected by their G418 resistance phenotype. The best ratio of homologous to nonhomologous recombination events was about 1:80. Analyses of the functional neo genes in various independent cell lines establish that simple crossovers (single and double) generated the wild-type neo genes.

MeSH Terms
Animals Chromosomes/physiology Cricetinae DNA Restriction Enzymes Genes Humans Hybrid Cells/cytology Kanamycin Kinase Phosphotransferases/genetics Plasmids Recombination, Genetic
Chemicals
Phosphotransferases Kanamycin Kinase DNA Restriction Enzymes
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Song K Y
Center for Genetics, University of Illinois College of Medicine, Chicago 60612.
Schwartz F
Maeda N
Smithies O
Kucherlapati 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.84.19.6820
Published
1987-10-00
Pages
6820-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC299176
Subset
IM
Grants
NIGMS NIH HHS · GM20069 · United States
NIGMS NIH HHS · GM33943 · United States
NIGMS NIH HHS · GM36565 · United States
External Links
PubMed source DOI: 10.1073/pnas.84.19.6820 PMC Full Text View journal details
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