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Ectopic germline recombination activity of the widely used Foxp3‐YFP‐Cre mouse: a case report
Author(s) -
Wu Dan,
Huang Qing,
Orban Paul C.,
Levings Megan K.
Publication year - 2020
Publication title -
immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.297
H-Index - 133
eISSN - 1365-2567
pISSN - 0019-2805
DOI - 10.1111/imm.13153
Subject(s) - biology , transgene , genotyping , genetics , germline , foxp3 , homologous recombination , cre recombinase , ectopic expression , microbiology and biotechnology , genetically modified mouse , gene , genotype , immune system
Summary Regulatory T (Treg) cell‐specific deletion of a gene of interest is a procedure widely used to study mechanisms controlling Treg development, homeostasis and function. Accordingly, several transgenic mouse lines have been generated that bear the Cre recombinase under control of the Foxp3 promoter either as a random transgene insertion or knocked into the endogenous Foxp3 locus, with the Foxp3 YFP‐Cre strain of mice being one of the most widely used. In an attempt to generate Treg cells that lacked expression of the insulin receptor (Insr), we crossed Foxp3 YFP‐Cre mice with Insr fl/fl mice. Using a conventional two‐band PCR genotyping method we found that offspring genotypes did not correspond to the expected Mendelian ratios. We therefore developed a quantitative PCR‐based genotyping method to investigate possible ectopic recombination outside the Treg lineage. With this method we found that ~50% of the F 1 ‐generation mice showed evidence of ectopic recombination and that ~10% of the F 2 ‐generation mice had germline Cre recombination activity leading to a high frequency of offspring with global Insr deletion. Use of the quantitative PCR genotyping method enabled accurate selection of mice without ectopic recombination and only the desired Treg cell‐specific Insr deletion. Our data highlight the need to use genotyping methods that allow for assessment of possible ectopic recombination driven by the Foxp3 YFP‐Cre allele, particularly when studying genes that are systemically expressed.