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Updated: Aug 12, 2026
Conditional Genetic Transsynaptic Tracing in the Embryonic Mouse Brain
Published on: December 22, 2014
Sequencing two Tyr::CreERT2 transgenic mouse lines
Zackie Aktary1,2,3, Andre Corvelo4, Camille Estrin1,2,3
1Institut Curie, INSERM U1021, Normal and Pathological Development of Melanocytes, PSL Research University, Orsay, France.
Abstract:
The Cre/loxP system is a powerful tool that has allowed the study of the effects of specific genes of interest in various biological settings. The Tyr::CreERT2 system allows for the targeted expression and activity of the Cre enzyme in the melanocyte lineage following treatment with tamoxifen, thus providing spatial and temporal control of the expression of specific target genes. Two independent transgenic mouse models, each containing a Tyr::CreERT2 transgene, have been generated and are widely used to study melanocyte transformation. In this study, we performed whole genome sequencing (WGS) on genomic DNA from the two Tyr::CreERT2 mouse models and identified their sites of integration in the C57BL/6 genome. Based on these results, we designed PCR primers to accurately, and efficiently, genotype transgenic mice. Finally, we discussed some of the advantages of each transgenic mouse model.
Insights
Researchers identified the integration sites of two Tyr::CreERT2 mouse models using whole genome sequencing. This information enables precise genotyping for studying melanocyte transformation.
Area of Science:
- Genetics and Genomics
- Developmental Biology
- Cancer Biology
Background:
- The Cre/loxP system enables targeted gene manipulation in biological research.
- The Tyr::CreERT2 mouse model allows tamoxifen-inducible Cre recombinase activity in melanocytes.
- Studying melanocyte transformation often utilizes transgenic models for genetic control.
Purpose of the Study:
- To identify the genomic integration sites of two independent Tyr::CreERT2 transgenic mouse models.
- To develop accurate PCR-based genotyping methods for these models.
- To compare the advantages of the characterized Tyr::CreERT2 mouse models.
Main Methods:
- Whole genome sequencing (WGS) of genomic DNA from Tyr::CreERT2 mouse models.
- Bioinformatic analysis to pinpoint transgene integration sites within the C57BL/6 genome.
- Design and validation of PCR primers for efficient and accurate genotyping.
Main Results:
- The specific integration sites for both Tyr::CreERT2 transgenes were successfully identified.
- Validated PCR primers provide a reliable method for distinguishing transgenic mice.
- Characterization facilitates informed selection between the two available mouse models.
Conclusions:
- Precise knowledge of transgene integration sites is crucial for characterizing transgenic models.
- The developed genotyping strategy enhances the utility of Tyr::CreERT2 mice for melanocyte research.
- This study provides valuable resources for researchers investigating melanocyte development and transformation.
