B6.Cg-Pabpc1l^tm1.1Esel/J

Stock number: 029031
Product name: Epab^-
Research areas: Cell Biology Research, Developmental Biology Research, Reproductive Biology Research

Description

Epab KO mice lack exon 2 of the Pabpc1l gene, making them useful for studying oogenesis, folliculogenesis and female fertility.

Detailed description

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.

Development

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

Symbol: Pabpc1l^tm1.1Esel
Name: targeted mutation 1.1, Emre Seli
MGI accession ID: MGI:5437740
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Pabpc1l
Marker name: poly(A) binding protein, cytoplasmic 1-like
Chromosome: 2
Strain of origin: 129S1/SvImJ or C57BL/6J
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.