DOI: 10.1101/gad.2.11.1353. Homologous integration in mammalian cells without target gene selection.

Homologous integration in mammalian cells without target gene selection.

10.1101/gad.2.11.1353

Crossref journal-article

Cold Spring Harbor Laboratory

Genes & Development (246)

Abstract

Homologous integrations into a nonselectable target locus have been highly enriched for following DNA transfections into mammalian cells. The target gene, the SV40 early region in COS1 cells, provides transcription signals to activate a defective selectable marker, the gpt gene. We find that nearly half of the selected clones have integrated the gpt gene at the homologous sequence in the COS1 genome. This is an estimated 100-fold enrichment for homologous events compared with transfections in which the gpt gene is transcriptionally active. As shown for yeast integration events, a double-strand break at a position of homology between the transfected DNA and the genomic target is necessary to achieve a high frequency of homologous integrations. Furthermore, the arrangement of sequences at the integration site includes a repair of the double-strand gap, which was present on the transfected DNA, suggesting that similarities exist between yeast and mammalian integrations. The experimental design, in which a defective marker is activated following a homologous integration, may have general applications for gene targeting in mammalian cells.

Bibliography

Jasin, M., & Berg, P. (1988). Homologous integration in mammalian cells without target gene selection. Genes & Development, 2(11), 1353â1363.

Authors 2

M Jasin (first)
P Berg (additional)

References 22 Referenced 123

{'key': '2021111319485304000_2.11.1353.1', 'first-page': '3246', 'article-title': 'Recombination of homologous DNA fragments transfected into mammalian cells occurs predominantly by terminal pairing.', 'volume': '6', 'year': '1986', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / Recombination of homologous DNA fragments transfected into mammalian cells occurs predominantly by terminal pairing. (1986)
{'key': '2021111319485304000_2.11.1353.2', 'first-page': '2520', 'article-title': 'Intramolecular recombination between transfected repeated sequences in mammalian cells is nonconservative.', 'volume': '6', 'year': '1986', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / Intramolecular recombination between transfected repeated sequences in mammalian cells is nonconservative. (1986)
10.1038/330576a0
10.1016/0092-8674(81)90282-8
10.1073/pnas.77.7.3898
10.1016/0092-8674(85)90242-9
{'key': '2021111319485304000_2.11.1353.7', 'first-page': '1020', 'article-title': 'Model for homologous recombination during transfer of DNA into mouse L cells: Role for DNA ends in the recombination process.', 'volume': '4', 'year': '1984', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / Model for homologous recombination during transfer of DNA into mouse L cells: Role for DNA ends in the recombination process. (1984)
10.1073/pnas.82.5.1391
Maniatis, T., E.F. Fritsch, and J. Sambrook. 1982. Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
10.1126/science.6251549
10.1073/pnas.78.4.2072
10.1073/pnas.80.14.4417
{'key': '2021111319485304000_2.11.1353.13', 'first-page': '2695', 'article-title': 'Termination-reinitiation occurs in the translation of mammalian cell mRNAs.', 'volume': '6', 'year': '1986', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / Termination-reinitiation occurs in the translation of mammalian cell mRNAs. (1986)
10.1073/pnas.76.10.4951
10.1101/SQB.1984.049.01.020
10.1038/317230a0
10.1073/pnas.84.19.6820
10.1016/0022-2836(75)90217-X
10.1016/0092-8674(83)90331-8
10.1016/0092-8674(87)90646-5
10.1016/0092-8674(86)90463-0
10.1016/0092-8674(84)90048-5

Dates

Type	When
Created	18 years, 2 months ago (June 5, 2007, 5:15 p.m.)
Deposited	3 years, 9 months ago (Nov. 13, 2021, 10:50 p.m.)
Indexed	1 month, 1 week ago (July 14, 2025, 11:47 p.m.)
Issued	36 years, 9 months ago (Nov. 1, 1988)
Published	36 years, 9 months ago (Nov. 1, 1988)
Published Online	36 years, 9 months ago (Nov. 1, 1988)
Published Print	36 years, 9 months ago (Nov. 1, 1988)

Funders 0

None

BibTeX

@article{Jasin_1988, title={Homologous integration in mammalian cells without target gene selection.}, volume={2}, ISSN={1549-5477}, url={http://dx.doi.org/10.1101/gad.2.11.1353}, DOI={10.1101/gad.2.11.1353}, number={11}, journal={Genes & Development}, publisher={Cold Spring Harbor Laboratory}, author={Jasin, M and Berg, P}, year={1988}, month=nov, pages={1353–1363} }

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