Epab KO mice lack exon 2 of the Pabpc1l gene, making them useful for studying oogenesis, folliculogenesis and female fertility.
Emre Seli, Yale Unversity School of Medicine
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Null/Knockout) | Pabpc1l | poly(A) binding protein, cytoplasmic 1-like |
Epab KO mice lack exon 2 of the poly(A) binding protein, cytoplasmic 1-like Pabpc1l gene, abolishing gene function. Exon 2 encodes a portion of the first RNA-recognition motif (RRM)1 domain and removal of it results in a premature stop codon and the production of a EPAB protein with truncated RRM1, and lacking the remaining three RRMs and poly(A)-binding (PAB) C-terminal domain.
EPAB is required for the regulation of maternal gene expression during oocyte development and maturation, fertilization, and early embryo development until zygotic genome activation. Homozygous Epab-/- males and heterozygous Epab+/- mice are viable and fertile, while homozygous Epab-/- females are viable but infertile. Homozygous Epab-/- females cannot generate embryos or mature oocytes in vivo or in vitro. Oocytes from these females failed to achieve translational activation of maternally-stored mRNAs upon stimulation of oocyte maturation. Spindle formation and chromosome alignment during meiotic divisions are impaired in Epab-/- oocytes. Microinjection of Epab mRNA into germinal vesicle stage Epab-/- oocytes did not rescue maturation, while microinjection of Epab mRNA into preantral follicle enclosed oocytes does.
A targeting vector was designed by Drs. Isaac Sasson, Maria D. Lalioti and Emre Seli (Yale University School of Medicine) to delete exon 2 of the poly(A) binding protein, cytoplasmic 1-like (Pabpc1l) gene. The insert of the targeting vector included a neomycin resistance (neo) cassette flanked by loxP sites and by Pabpc1l sequences corresponding to introns 1 and 2 of the gene. The construct was electroporated into 129S1/SvImJ and C57BL/6J-derived S1B6A embryonic stem (ES) cells. Correctly targeted ES cells were transfected with a Cre-expressing transgene to remove the neo cassette. These ES cells were injected into C57BL/6 blastocysts and the resulting chimeric males were bred to C57BL/6 females. The resulting Epab KO mice were bred to C57BL/6J mice for 40 generations. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).
Allele Name | targeted mutation 1.1, Emre Seli |
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Allele Type | Targeted (Null/Knockout) |
Allele Synonym(s) | |
Gene Symbol and Name | Pabpc1l, poly(A) binding protein, cytoplasmic 1-like |
Gene Synonym(s) | |
Strain of Origin | 129S1/SvImJ or C57BL/6J |
Chromosome | 2 |
Molecular Note | A floxed neo cassette replaced exon 2. Cre-mediated recombination removed the neo cassette. RT-PCR confirmed the absence of protein expression in ovaries. |
When maintaining a live colony, heterozygous females may be bred to homozygous males. Homozygous KO female mice are infertile.
When using the Epab- mouse strain in a publication, please cite the originating article(s) and include JAX stock #029031 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous for Pabpc1l<tm1.1Esel> |
Frozen Mouse Embryo | B6.Cg-Pabpc1l<tm1.1Esel>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Pabpc1l<tm1.1Esel>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Pabpc1l<tm1.1Esel>/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6.Cg-Pabpc1l<tm1.1Esel>/J Frozen Embryo | $3373.50 |
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