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Generation of a germ cell nuclear factor conditional allele in mice
Author(s) -
Lan ZiJian,
Xu Xueping,
Cooney Austin J.
Publication year - 2003
Publication title -
genesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.093
H-Index - 110
eISSN - 1526-968X
pISSN - 1526-954X
DOI - 10.1002/gene.10239
Subject(s) - germline , biology , exon , allele , germ cell , microbiology and biotechnology , transgene , cre recombinase , genetics , genetically modified mouse , gene
Summary: Two recombination steps in embryonic stem (ES) cells were adopted to generate a floxed Germ Cell Nuclear Factor ( GCNF ) allele. First, a targeting vector containing a loxP site upstream of exon 4, encoding the DNA binding domain (DBD), and a floxed NeoTK double selection cassette downstream of exon 4 was integrated into the GCNF locus by homologous recombination. Second, a Cre‐expressing vector was transiently introduced to remove the floxed NeoTK cassette via site‐specific recombination. Heterogeneous ES cell populations were found in a single colony after Cre transfection and were separated using an ES cell re‐pick step. Floxed GCNF mice were generated and had normal GCNF expression in the adult gonads. Using the Msx2Cre transgenic mice, the floxed GCNF can be completely deleted in the female germline. Taken together, the floxed GCNF mice were successfully generated and female germline deletion of the floxed GCNF allele was achieved using Msx2Cre mice. genesis 37:172–179, 2003. © 2003 Wiley‐Liss, Inc.