title: 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.
authors.affiliation: Department of Bioscience, Tokyo University of Agriculture, Tokyo 156-8502, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Hara
authors.forename: Takashi
authors.initials: T
authors.lastname: Takano
authors.forename: Mio
authors.initials: M
authors.lastname: Ogata
authors.forename: Reina
authors.initials: R
authors.lastname: Yamakami
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Sato
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Kono
authors.forename: Yayoi
authors.initials: Y
authors.lastname: Obata
mesh_terms: D000818:Animals; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics / Q000378:metabolism; D000090205:DNA Methyltransferase 3A; D005260:Female; D018014:Gene Transfer Techniques / Q000662:veterinary; D017930:Genes, Reporter; D005854:Germ Cells / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D041681:NIH 3T3 Cells; D009866:Oogenesis / Q000235:genetics; D010442:Peptide Chain Initiation, Translational / Q000235:genetics; D010446:Peptide Fragments / Q000235:genetics* / Q000378:metabolism; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D019076:Transgenes; D000097573:Red Fluorescent Protein
chemical_list: D004248:DNA (Cytosine-5-)-Methyltransferases; D000090205:DNA Methyltransferase 3A; D008164:Luminescent Proteins; D010446:Peptide Fragments; D000097573:Red Fluorescent Protein