These Sec24c genetrap mutant mice incorporate a lacZ reporter and carry a floxed exon 3. They are useful in studies of embryonic development and endoplasmic reticulem (ER)-Golgi vesicular trafficking.
David Ginsburg, University of Michigan
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.
These mice carry the Sec24c-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 2 of the gene and and additional loxP site in intron 3. When FLP recombinase is introduced, an allele carrying a floxed exon 3, but lacking both the lacZ and neomycin features, can be produced (see Stock No. 024867). Cre excision of the floxed segment results in a frameshift mutation and early stop in exon 4 producing a null allele (see Stock No. 024868).
Heterozygous Sec24c-GT mice are healthy and appear normal, but homozygotes exhibit embryonic lethality due to the presence of the gene trap cassette.
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). This gene trapped trapped allele of Sec24c 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 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)-Tyrc-2J/J (see Stock No. 000058) to achieve germline transmission. This strain was backcrossed to C57BL/6J for 6 generations and the Tyrc-2J allele was bred out of the line by the donating laboratory.
|Expressed Gene||lacZ, beta-galactosidase, E. coli|
|Site of Expression|
|Allele Name||targeted mutation 1a, Wellcome Trust Sanger Institute|
|Allele Type||Targeted (Conditional ready (e.g. floxed), Reporter, Null/Knockout)|
|Gene Symbol and Name||Sec24c, Sec24 related gene family, member C (S. cerevisiae)|
|Expressed Gene||lacZ, beta-galactosidase, E. coli|
|Strain of Origin||C57BL/6N|
|General Note||Cell line EPD0241_2_A11 was successfully used to make chimeric mice. Germline transmission was accomplished. J:232065|
|Molecular Note||The L1L2_Pgk_P cassette was inserted at position 20679037 of Chromosome 14 upstream of the critical exon(s) (Build GRCm38). 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 PGK 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 20679926. 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|
Heterozygotes are viable and fertile. Homozygotes die during embryonic development due to the presence of the genetrap cassette.
When using the B6J.B6N(Cg)-Sec24ctm1a(EUCOMM)Wtsi/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #024866 in your Materials and Methods section.
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