Cited by 944Animal Genetics and ReproductionCRISPR and Genetic EngineeringPluripotent Stem Cells Research

Generation of mice carrying a mutant apolipoprotein E gene inactivated by gene targeting in embryonic stem cells.

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Piedrahita, J A, Zhang, S H, Hagaman, J R, Oliver, P M, Maeda, N·1992·Proceedings of the National Academy of Sciences

Abstract

We have inactivated the endogenous apolipoprotein E (apoE) gene by using gene targeting in mouse embryonic stem (ES) cells. Two targeting plasmids were used, pJPB63 and pNMC109, both containing a neomycin-resistance gene that replaces a part of the apoE gene and disrupts its structure. ES cell colonies targeted after electroporation with plasmid pJPB63 were identified by the polymerase chain reaction (PCR) followed by genomic Southern analysis. Of 648 G418-resistant colonies analyzed, 9 gave a positive signal after PCR amplification, and 5 of them were confirmed as targeted by Southern blot analysis. The second plasmid, pNMC109, contains the negatively selectable thymidine kinase gene in addition to the neomycin-resistance gene. After electroporation with this plasmid, 177 colonies resistant both to G418 and ganciclovir were analyzed; 39 contained a disrupted apoE gene as determined by Southern blotting. Chimeric mice were generated by blastocyst injection with 6 of the targeted lines. One of the lines gave strong chimeras, three of which transmitted the disrupted apoE gene to their progeny. Mice homozygous for the disrupted gene were produced from the heterozygotes; they appear healthy, even though they have no apolipoprotein E in their plasma.

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Crossref · 10.1073/pnas.89.10.4471

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Authors
Piedrahita, J. A., Zhang, S. H., Hagaman, J. R., Oliver, P. M., & Maeda, N.
Affiliations
University of North Carolina at Chapel Hill (US)
Venue
Proceedings of the National Academy of Sciences
Published
1992-05-15
Volume
89
Issue
10
Pages
4471-4475
ISSN
  • 0027-8424 (ISSN-L)
  • 1091-6490
OpenAlex
W2062108627
Cited by
944
References
38
Citations by year
2026: 12 · 2025: 24 · 2024: 21 · 2023: 18 · 2022: 29 · 2021: 28 · 2020: 26 · 2019: 32 · 2018: 29 · 2017: 42 · 2016: 31 · 2015: 23 · 2014: 28 · 2013: 34 · 2012: 31
FWCI
10.25
Type
journal article
Language
en
Open access
Locations
  • Proceedings of the National Academy of Sciences (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • Carolina Digital Repository (University of North Carolina at Chapel Hill) (submittedVersion)
  • UNC Libraries
Funders
  • National Institutes of Health
  • National Heart, Lung, and Blood Institute (HL 42630)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Animal Genetics and Reproduction
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
Keywords
apolipoprotein E, gene targeting, mouse embryonic stem cells, knockout mice, neomycin resistance, thymidine kinase, chimeric mice, blastocyst injection, gene inactivation
MeSH terms
Animals, Apolipoproteins E, Base Sequence, Cell Line, Chimera, DNA, Embryo, Mammalian, Mice, Mutant Strains, Molecular Sequence Data, Oligodeoxyribonucleotides, Plasmids, Recombination, Genetic, Stem Cells, Blotting, Southern, Restriction Mapping, Polymerase Chain Reaction, Mice

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