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Cited by 316

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Conditional gene targeted deletion by Cre recombinase demonstrates the requirement for the double-strand break repair Mre11 protein in murine embryonic stem cells

Open access
Xiao, Y·1997·Nucleic Acids Research

Abstract

Repair of DNA damage resulting in double-strand breaks (DSBs) is controlled by gene products executing homologous recombination or end-joining pathways. The MRE11 gene has previously been implicated in DSB repair in the yeast Saccharomyces cerevisiae . Here we have developed a methodology to study the roles of the murine Mre11 homolog in pluripotent embryonic stem cells. Using a gene targeting approach, a triple LoxP site cassette was inserted into a region of MRE11 genomic DNA flanking conserved phosphodiesterase motifs. The addition of Cre recombinase activity promotes deletions of three types that can be scored. We find that deletion at phosphodiesterase motif III encoded in the N-terminus of Mre11 is acheived in the presence of a wild-type MRE11 allele. However, when the wild-type MRE11 allele is inactivated by gene targeted insertion of a neo marker, only Cre recombination events that allow expression of wild-type Mre11 protein are observed. Therefore, Mre11 is required for normal cell proliferation. This methodology introduces a means to study important regions of essential genes in cell culture models.

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Metadata

Authors
Xiao, Y.
Venue
Nucleic Acids Research
Published
1997-08-01
Volume
25
Issue
15
Pages
2985-2991
ISSN
  • 0305-1048 (ISSN-L)
  • 1362-4962
OpenAlex
W2157935839
Cited by
316
References
31
FWCI
3.04
Type
journal article
Language
en
Open access
green

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CoChat. (2026). Citation check: Conditional gene targeted deletion by Cre recombinase demonstrates the requirement for the double-strand break repair Mre11 protein in murine embryonic stem cells. https://app.cochat.ai/papers/10.1093/nar/25.15.2985
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Xiao, Y. (1997). Conditional gene targeted deletion by Cre recombinase demonstrates the requirement for the double-strand break repair Mre11 protein in murine embryonic stem cells. Nucleic Acids Research. https://doi.org/10.1093/nar/25.15.2985

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