This page contains recommended breeding strategies and protocols. If you have any questions, please contact us at firstname.lastname@example.org.
Germline Transmission Testing (ES cell)
- MMRRC and MBP Germline Transmission Testing Policy
- C57BL/6N Non-Agouti Protocol (JM8 sublines, VGB6)
- C57BL/6N Agouti Protocol (JM8A sublines)
- BayGenomics and SIGTR Gene Trap Protocol
GENSAT Breeding Recommendations
In general, GENSAT BAC transgenic mouse lines should be utilized in the hemizygous state. Breeding of the BAC transgene to a homozygous state carries an increased risk of generating an unwanted phenotype for two reasons. First, the transgene inserts at random into the genome. Breeding the transgene to homozygosity results in mice that carry this insertion on both chromosomes, increasing the probability that the insertion will result in an observable phenotype. Second, BAC transgenes may carry passenger genes that are also expressed from the BAC vector. Breeding to homozygosity doubles the number of these genes carried in the transgenic mouse, increasing the possibility that a phenotype will result as a consequence of increased gene dosage. In our experience, observable phenotypes are rare in BAC transgenic mice when carried in the hemizygous state.
Reduced expression may be a concern over time for some Cre and EGFP lines maintained in live colonies. There is a publication with regards to variable expression for Cre lines, specifically a decrease over time, while still detecting it at the genomic level: http://www.plosone.org/article/info:doi/10.1371/journal.pone.0001013. There have also been studies that show GFP expression can deteriorate over time: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5050239/
Mixed Background Strain Recommendations
For strains on a mixed background (such as mixed B6;129), we recommend using wildtype animals generated from heterozygous matings as controls. If homozygous mice are needed, we recommend maintaining your colony with heterozygous to heterozygous matings, as this will generate both homozygous and wildtype mice.