B6(Cg)-Sec24d^{tm1c(EUCOMM)Hmgu}/DgiJ

Stock number: 025889
Research areas: Cell Biology Research, Research Tools

Description

Exons 5 and 6 of the Sec24d gene are flanked by loxP sites in this conditional Sec24d-FL 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. Sec24d (Sec24 related gene family, member D (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 SEC24D die prior to the blastocyst stage of embryonic development.

Exons 5 and 6 of the Sec24d gene are flanked by loxP sites in this conditional mutant strain. Heterozygous and homozygous Sec24c-FL mice are healthy, appear normal and are fertile. Cre excision of the floxed segment is believed to result in the production of a truncated protein product which may be subject to non-sense-mediated decay, producing a null allele (see Stock No. 025749).

Development

Embryonic stem (ES) cell clone HEPD0753_7-H04 (EUCOMM ID: LOC383951) was created as part of the Knockout Mouse Project (KOMP). The original gene trapped allele of Sec24d 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 Sec24d-GT (tm1a(EUCOMM)Hmgu) allele. C57BL/6N-A^tm1Brd-derived JM8A3.N1 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 Actin-Flp recombinase (see Stock No. 005703), leaving exons 5 and 6 flanked by loxP sites. Mice were backcrossed to C57BL/6J for one generation to remove the FLP transgene. The Tyr^c-2J allele was bred out of the line. The donating laboratory observed agouti and black animals in their colony.

Allele

Symbol: Sec24d^{tm1c(EUCOMM)Hmgu}
Name: targeted mutation 1c, Helmholtz Zentrum Muenchen GmbH
MGI accession ID: MGI:5582296
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Synonyms: Sec24d-FL
Marker symbol: Sec24d
Marker name: Sec24 related gene family, member D (S. cerevisiae)
Marker synonyms: 2310020L09Rik; CLCRP2; LOC383951
Chromosome: 3
Strain of origin: C57BL/6N-A^tm1Brd
Molecular note: The L1L2_Bact_P cassette was inserted at position 123086679 of Chromosome 3 upstream of the critical exon(s) (Build GRCm39). 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 human beta-actin promoter, SV40 polyA, a second FRT site and a second loxP site. A third loxP site is inserted downstream of the targeted exon(s) at position 123088075. The critical exon(s) is/are thus flanked by loxP sites. The lacZ and neomycin segments were excised with germline Actin-Flp recombinase, leaving exons 5 and 6 flanked by loxP sites. Further information on targeting strategies used for this and other IKMC alleles can be found at http://www.informatics.jax.org/mgihome/nomen/IKMC_schematics.shtml.