Disruption of the proto-oncogene int-2 in mouse embryo-derived stem cells: A general strategy for targeting mutations to non-selectable genes
Abstract
Gene targeting--homologous recombination of DNA sequences residing in the chromosome with newly introduced DNA sequences--in mouse embryo-derived stem cells promises to provide a means to generate mice of any desired genotype. We describe a positive nd negative selection procedure that enriches 2,000-fold for those cells that contain a targeted mutation. The procedure was applied to the isolation of hprt- and int-2- mutants, but it should be applicable to any gene.
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Due to the extremely low frequency of homologous recombination, this technique cannot be used for human gene therapy.
“However, due to the extremely low frequency of HR, this technique cannot be used for human gene therapy.”
From Newer Gene Editing Technologies toward HIV Gene Therapy (2013) · cites this paper as [15]
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“While a variety of gene targeting approaches have been employed for hundreds of mouse loci, the majority of Ahr alleles were modified using the positive negative selection approach pioneered by Capecchi and colleagues (Mansour et al. 1988).”
Method
- Authors
- Mansour, S. L., Thomas, K. R., & Capecchi, M. R.
- Affiliations
- Howard Hughes Medical Institute (US)
- Venue
- Nature
- Published
- 1988-11
- Volume
- 336
- Issue
- 6197
- Pages
- 348-352
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- OpenAlex
- W1983237714
- Cited by
- 1,683
- References
- 15
- Citations by year
- 2026: 2 · 2025: 18 · 2024: 9 · 2023: 14 · 2022: 14 · 2021: 16 · 2020: 12 · 2019: 18 · 2018: 15 · 2017: 15 · 2016: 26 · 2015: 24 · 2014: 26 · 2013: 29 · 2012: 32
- FWCI
- 21.48
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Animal Genetics and Reproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Keywords
- gene targeting, homologous recombination, positive and negative selection, knockout mice
- MeSH terms
- Animals, Clone Cells, Mice, Transgenic, Mutation, Proto-Oncogenes, Stem Cells, Transfection, Restriction Mapping, Mice
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