B6J.B6N(Cg)-Sec24d^{tm1a(EUCOMM)Hmgu}/DgiJ

Stock number: 025748
Research areas: Cell Biology Research, Developmental Biology Research, Research Tools

Description

These Sec24d genetrap mutant mice (Sec24d-GT) incorporate a lacZ reporter and carry floxed exons 5 and 6. They 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 during embryonic development.

These mice carry the Sec24d-GT genetrap allele that incorporates an FRT-En2 splice acceptor (SA)-IRES-lacZ-SV40 polyadenylation signal (pA)-loxP-PGK-Neo-SV40 polyadenylation signal (pA)-FRT-loxP sequence in intron 4 of the gene and and additional loxP site in intron 6. When FLP recombinase is introduced, an allele carrying a floxed exons 5 and 6, but lacking both the lacZ and neomycin features, can be produced (see Stock No. 025889). Cre excision of the floxed segment is anticipated to result in a transcript producing truncated protein subject to non-sense-mediated decay (see Stock No. 025749).

Heterozygous Sec24d-GT mice are healthy and appear normal, but homozygotes are likely to be embryonic lethal. Functionality of the lacZ component has not been tested and assays have not been done to determine whether a predicted truncated protein is produced.

Development

Embryonic stem (ES) cell clone HEPD0753_7_H04 (EUCOMM ID: LOC383951) was created by the European Conditional Mouse Mutagenesis Program (EUCOMM) as part of the Knockout Mouse Project (KOMP). This gene trapped allele of Sec24d incorporates an FRT-En2 splice acceptor (SA)-IRES-lacZ-SV40 polyadenylation signal (pA)-loxP-PGK-Neo-SV40 polyadenylation signal (pA)-FRT-loxP sequence in intron 4 of the gene and an additional loxP site in intron 6 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. This strain was backcrossed to C57BL/6J for four generations and the Tyr^c-2J allele was bred out of the line by the donating laboratory. The donating lab observed agouti and black animals in their colony.

Allele

Symbol: Sec24d^{tm1a(EUCOMM)Hmgu}
Name: targeted mutation 1a, Helmholtz Zentrum Muenchen GmbH
MGI accession ID: MGI:5287846
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter; Null/Knockout
Marker symbol: Sec24d
Marker name: Sec24 related gene family, member D (S. cerevisiae)
Chromosome: 3
Strain of origin: C57BL/6N-A^tm1Brd
Expressed genes: lacZ,beta-galactosidase,E. coli; neo,neomycin resistance,bacterial
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 a 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. A "conditional ready" (floxed) allele can be created by flp recombinase expression in mice carrying this allele. Subsequent cre expression results in a knockout mouse. If cre expression occurs without flp expression, a reporter knockout mouse will be created. 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
General note: Homozygotes are expected to exhibit embryonic lethality.