Establishment of a Conditional Transgenic System Using the 2A Peptide in the Female Mouse Germline

Abstract

Transgenic mice are essential research tools in developmental biology studies. The 2A peptide allows multiple genes to be expressed simultaneously at comparable levels in somatic cells, but there are no reports of it being used successfully in germ cells. We constructed a Cre/loxP-based conditional vector containing the 2A peptide to significantly enhance the expression of a reporter and target gene from a constitutive promoter in oocytes. Mice with a transgene insertion containing the chicken β-actin promoter, floxed EGFP-polyA cassette, mCherry reporter, 2A peptide and target gene DNA methyltransferase 3A2 (Dnmt3a2) were crossed with TNAP- or Vasa-Cre mice to produce offspring, in which mCherry and DNMT3A2 proteins were highly expressed in oocytes upon Cre-mediated removal of EGFP-polyA. This novel transgenic mouse line based on the 2A expression system can serve as a useful tool for examining gene function during oogenesis.

Ectopic expression of DNA methyltransferases DNMT3A2 and DNMT3L leads to aberrant hypermethylation and postnatal lethality in mice

“…The previously generated 2lox mice, harboring floxed alleles were used in this study (Hara, Takano, Fujikawa, et al, ; Hara, Takano, Ogata, et al, ). For the production of DNMTs‐Tg mice (1lox mice), Vasa ‐Cre female mice, having FVB genetic background (purchase from the Jackson Laboratory, Bar Harbor, ME), were crossed with 2lox male mice.…”

“…The previously generated 2lox mice, harboring floxed alleles were used in this study (Hara, Takano, Fujikawa, et al, 2014;Hara, Takano, Ogata, et al, 2014). For the production of DNMTs-Tg mice (1lox mice),…”