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Conditional gene mutagenesis in B-lineage cells
1The CBR Institute for Biomedical Research, Harvard Medical School, Boston, MA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|May 18, 2004
Summary
The Cre/loxP system enables precise gene function studies in mice. Updated protocols guide conditional gene targeting, embryonic stem cell manipulation, and B-cell development research.
Area of Science:
- Molecular Biology
- Genetics
- Immunology
Background:
- The Cre/loxP recombination system is a powerful tool for genetic engineering.
- It allows for cell-type, stage-specific, and inducible control of gene function in vivo.
- Its application in mice has revolutionized the study of gene function.
Purpose of the Study:
- To provide updated protocols for utilizing the Cre/loxP system.
- To facilitate conditional gene targeting and embryonic stem cell manipulation.
- To aid in the assessment of gene function during B-cell development.
Main Methods:
- Vector construction for conditional gene targeting.
- In vitro manipulation of embryonic stem (ES) cells.
- Identification of successfully targeted ES colonies.
- Generation of single-copy, Cre-dependent knock-in transgenes.
Main Results:
- Updated protocols for Cre/loxP system implementation.
- Guidelines for generating Cre-regulated transgenes.
- A comprehensive list of Cre strains for B-cell development studies.
Conclusions:
- The Cre/loxP system is essential for in vivo gene function analysis.
- Updated protocols enhance the efficiency of gene targeting and ES cell manipulation.
- This resource aids researchers in studying gene function during B-cell development.