Vps34f/f floxed mice are used to conditionally delete Pik3c3, making them useful for studies of autophagy and exocytosis.
Jianhua Zhang, University of Alabama at Birmingham
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Conditional ready (e.g. floxed)) | Pik3c3 | phosphatidylinositol 3-kinase catalytic subunit type 3 |
These Vps34f/f mutant mice possess loxP sites flanking exon 4 of the phosphoinositide-3-kinase, class 3 (Pik3c3) gene. Also known as Vsp34, Pik3c3 generates phosphatidylinositol 3-phosphate, a regulator of membrane trafficking such as autophagy and endocytosis. Mice that are homozygous for this allele are viable and fertile. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have exon 4 deleted in the cre-expressing tissues.
For example, when bred to B6.Cg-Tg(Alb-cre)21Mgn/J mice (Stock No. 003574) expressing cre recombinase in liver, resulting mice develop hepatomegaly and hepatic steatosis, with impaired protein turnover. These double mutant mice exhibit decreased survival, with 50% lethality in the first 2 months of life.
For example, when bred to B6.FVB(129S4)-Tg(Ckmm-cre)5Khn/J mice (Stock No. 006475) expressing cre recombinase in heart, mice exhibit cardiomegaly and decreased contractility. These double mutant mice die between 5 and 13 weeks of age.
A targeting vector was designed to insert, from 5' to 3', a frt-site, a β-galactosidase cassette, a neomycin resistance cassette, a second frt-site, and a loxP site upstream of exon 4 of the phosphoinositide-3-kinase, class 3 (Pik3c3) gene. A second loxP site was also inserted just downstream of exon 4 of the targeted gene. The construct was electroporated into C57BL/6N-derived JM8.F6 embryonic stem (ES) cells, and correctly targeted ES cells were injected into blastocysts. Resulting chimeric males were bred to C57BL/6 females, and their offspring were bred to FLP-expressing mice to delete the β-gal and neo cassette. These conditional Vps34f/f mice were maintained on a mixed background. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6NJ (Stock No. 005304) for at least one generation to establish the colony.
Allele Name | targeted mutation 1c, Wellcome Trust Sanger Institute |
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Allele Type | Targeted (Conditional ready (e.g. floxed)) |
Allele Synonym(s) | Vps34f |
Gene Symbol and Name | Pik3c3, phosphatidylinositol 3-kinase catalytic subunit type 3 |
Gene Synonym(s) | |
Strain of Origin | C57BL/6N |
Chromosome | 18 |
General Note | Cell line EPD0103_4_A07 was used to make chimeric mice. Germline transmission was accomplished and subsequenct recombination established the allele. J:182628 |
Molecular Note | An FRT-flanked promoterless cassette, which contains a splice acceptor, beta-geo and polyadenylation sequence, with a 3' loxP site was inserted upstream of exon 4. An additional loxP site was inserted downstream of exon 4. Flp-mediated recombination removed the promoterless cassette and left exon 4 floxed. |
Mutations Made By | Jianhua Zhang, University of Alabama at Birmingham |
When maintaining a live colony, homozygous mice may be bred together.
When using the Vsp34f mouse strain in a publication, please cite the originating article(s) and include JAX stock #019081 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Heterozygous or wildtype for Pik3c3<tm1c(EUCOMM)Wtsi> |
Frozen Mouse Embryo | STOCK Pik3c3<tm1c(EUCOMM)Wtsi>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | STOCK Pik3c3<tm1c(EUCOMM)Wtsi>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | STOCK Pik3c3<tm1c(EUCOMM)Wtsi>/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | STOCK Pik3c3<tm1c(EUCOMM)Wtsi>/J Frozen Embryo | $3373.50 |
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