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Cre-mediated somatic site-specific recombination in mice
1Division of Molecular Genetics, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.
Abstract:
Conditional mutant mice equipped with heterologous recombination systems (Cre/lox or Flp/frt) are promising for studying tissue-specific gene function and for designing better models of human diseases. The utility of these mice depends on the cell target specificity, on the efficiency and on the control over timing of gene (in)activation. We have explored the utility of adenoviral vectors and transgenic mice expressing Cre under the control of tissue-specific promoters to achieve Cre/lox-mediated somatic recombination of the LacZ reporter gene, using a newly generated flox LacZ mouse strain. When adeno Cre viruses were administered via different routes, recombination and expression of LacZ was detected in a wide range of tissues. Whereas in liverbeta-galactosidase activity was quickly lost by turnover of expressing cells, even though the recombined allele was retained,beta-galactosidase in other tissues persisted for many months. Our data indicate that the flox LacZ transgenic line can be utilized effectively to monitor the level and functionality of Cre protein produced upon infection with adeno Cre virus or upon crossbreeding with different Cre transgenic lines.
Insights
Conditional mutant mice using Cre/lox systems are valuable for studying gene function and disease models. A new flox LacZ mouse strain effectively monitors Cre-mediated recombination across various tissues.
Area of Science:
- Genetics
- Molecular Biology
- Animal Models
Background:
- Conditional mutant mice with recombination systems (Cre/lox, Flp/frt) are crucial for tissue-specific gene function studies and disease modeling.
- The effectiveness of these systems relies on cell target specificity, efficiency, and temporal control of gene activity.
Purpose of the Study:
- To evaluate the utility of adenoviral vectors and transgenic mice expressing Cre for Cre/lox-mediated somatic recombination.
- To assess the functionality of a newly generated flox LacZ reporter mouse strain for monitoring Cre activity.
Main Methods:
- Administration of adenoviral Cre vectors via different routes in flox LacZ mice.
- Generation and utilization of transgenic mice expressing Cre under tissue-specific promoters.
- Monitoring LacZ reporter gene recombination and beta-galactosidase activity.
Main Results:
- Adeno-Cre virus administration resulted in LacZ recombination and expression in numerous tissues.
- Beta-galactosidase activity in the liver was transient due to cell turnover, while in other tissues it persisted for months.
- The flox LacZ strain effectively reported Cre protein level and functionality.
Conclusions:
- The flox LacZ mouse line is a versatile tool for assessing Cre recombinase activity from various sources.
- This model system aids in the study of gene function and the development of better disease models.