B6(Cg)-Sec24c^{tm1d(EUCOMM)Wtsi}/J

Stock number: 024868
Research areas: Cell Biology Research

Description

Exon 3 of the Sec24c gene has been deleted in this Sec24c-Null mutant strain. These mice are useful in studies of embryonic development and endoplasmic reticulem (ER)-Golgi vesicular trafficking.

Detailed description

SEC24 proteins are responsible for selectively recruiting cargo proteins from the endoplasmic reticulum (ER) into COPII vesicles that mediate transport to the Golgi. Sec24c (Sec24 related gene family, member C (S. cerevisiae)) is one of four mammalian Sec24 paralogs which are highly conserved at the C-terminus and variable in the N-terminal region. Mice completely lacking SEC24C die around embryonic day 7.

Exon 3 of the Sec24c gene has been deleted in this null allele. Homozygotes die by embryonic day 7-8.5 for undetermined reasons. Heterozygotes can be intercrossed with Sec24c-FL floxed mice (see Stock No. 024867), then further crossed with Cre strains to develop tissue specific knockouts. Cre strains utilized by the donating laboratory include: P48-Cre (pancreas-specific), villin-Cre (intestinal-epithelial-specific (see Stock No. 004586)), albumin-Cre (hepatocyte-specific (see Stock No. 003574)),SM22-Cre (smooth-muscle-specific (see Stock No. 04746)), and Meox2-Cre (ubiquitous expression beginning at embryonic day 5 (see Stock No. 003755)).

Development

Embryonic stem (ES) cell clone EPD0241-2-A11 (EUCOMM ID: 32981) was created by the Wellcome Trust Sanger Institute as part of the Knockout Mouse Project (KOMP). The original gene trapped allele of Sec24c incorporated an FRT-En2 splice acceptor (SA)-IRES-lacZ-SV40 polyadenylation signal (pA)-loxP-PGK-Neo-SV40 polyadenylation signal (pA)-FRT-loxP sequence in intron 2 of the gene and an additional loxP site in intron 3 to create the Sec24c-GT (tm1a(EUCOMM)Wtsi) allele. C57BL/6N-derived JM8.N4 embryonic stem (ES) cells were used to create the mutation. Chimeras were crossed with B6(Cg)-Tyr^c-2J/J (see Stock No. 000058) to achieve germline transmission. The lacZ and neomycin segments were excised with germline Flp recombinase (see Stock No. 005703), leaving exon 3 flanked by loxP sites. Then animals were crossed with a germline Cre strain (see Stock No. 003724) to delete exon 3. This strain was backcrossed to C57BL/6J for 3 generations by the donating laboratory. The Tyr^c-2J allele was bred out of the line.

Allele

Symbol: Sec24c^{tm1d(EUCOMM)Wtsi}
Name: targeted mutation 1d, Wellcome Trust Sanger Institute
MGI accession ID: MGI:5501471
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Sec24c
Marker name: Sec24 related gene family, member C (S. cerevisiae)
Chromosome: 14
Strain of origin: C57BL/6N
Molecular note: The L1L2_Pgk_P cassette was inserted at position 20679037 of Chromosome 14 upstream of exon 3. The cassette is composed of an FRT site followed by lacZ sequence and a loxP site. This first loxP site is followed by neomycin resistance gene under the control of the PGK promoter, SV40 polyA, a second FRT site and a second loxP site. A third loxP site is inserted downstream of exon 3 at position 20679926. A null allele was created by flp and cre recombinase expression in mice, resulting in the deletion of the lacZ sequences, the neomycin selection cassette and exon 3.
General note: Cell line EPD0241_2_A11 was successfully used to make chimeric mice. Germline transmission was accomplished. J:232065