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Efficient Cre-loxP-induced mitotic recombination in mouse embryonic stem cells
Pentao Liu1, Nancy A Jenkins, Neal G Copeland
1Mouse Cancer Genetics Program, Center for Cancer Research, National Cancer Institute at Frederick, Frederick, Maryland 21702, USA.
Nature Genetics
|December 12, 2001
Summary
Mitotic recombination can be induced in mouse embryonic stem cells using Cre/loxP technology. This method enables genetic mosaic analysis in mice, producing clones of homozygous mutant cells from heterozygous mothers.
Area of Science:
- Developmental Biology
- Genetics
- Molecular Biology
Background:
- FLP/FRT-induced mitotic recombination is a key technique for creating genetic mosaics in Drosophila.
- It allows for the functional analysis of recessive genes in heterozygous individuals.
Purpose of the Study:
- To demonstrate the reproducible induction of mitotic recombination in mouse embryonic stem (ES) cells.
- To assess the feasibility of genetic mosaic analysis in mice using Cre/loxP technology.
Main Methods:
- Utilized Cre/loxP technology for inducing mitotic recombination in mouse ES cells.
- Investigated recombination frequencies with single and multiple allelic loxP sites.
- Analyzed the timing of recombination (G2 phase) and subsequent X segregation during mitosis.
Main Results:
- Achieved reproducible induction of mitotic recombination in mouse ES cells with frequencies up to 5.0 x 10(-2).
- Observed that recombination predominantly occurs in the G2 phase, followed by X segregation.
- Confirmed that X segregation produces clones of homozygous mutant daughter cells.
Conclusions:
- Genetic mosaic analysis is feasible in mice using Cre/loxP-induced mitotic recombination.
- X segregation, crucial for mosaic analysis, is a conserved feature of mitosis across eukaryotes.
- This technique holds potential for studying gene function in specific chromosomal regions in mice.