Pfdn1 KO mice may be useful in studying the role of cytoskeletal function and remodeling on immune cell signaling and axon guidance.
Dr. Wasif Khan, University of Miami
Pfdn1 KO mice have a gene trap cassette inserted into intron 3 of the prefoldin 1 (Pfdn1) gene. PFDN1 encodes the first subunit of a molecular chaperone complex that binds and stabilizes newly synthesized actins and cytoskeletal proteins, allowing them to fold correctly. Heterozygous mice are viable and fertile, while homozygous mice die before five weeks of age due to neuromuscular dysfunction and physical wasting. Insertion of the gene trap sequence results in deletion of the coiled-coil domain required for interactions between Prefoldin and peptide substrates. These mice exhibit defects in cytoskeletal function such as ciliary dyskinesia, neuronal loss, and defects in B and T cell development and function. B and T cell maturation is impaired at the transitions from pre-pro-B to pre-B cells in the bone marrow and from CD4-CD8- double-negative to CD4+CD8+ double-positive T cells in the thymus.
Gene trap ES cell line GTR1.3, generated in 129/Sv-derived embryonic stem cells, was used to produce these Pfdn1 KO mice. The gene trap vector contains (from 5’ to 3’) a puromycin selection cassette, an internal ribosomal entry site (IRES), a β-galactosidase reporter (lacZ) with a polyadenylation signal, an RNA polymerase 2 gene (Pol2), and a neomycin selection sequence. This gene trap was inserted into intron 3 of the prefoldin 1 (Pfdn1) gene. These ES cells were injected into recipient blastocysts and chimeric males were bred with C57BL/6J females. Pfdn1 KO mice were backcrossed to C57BL/6J mice for at least six generations. Upon arrival at The Jackson Laboratory, 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||gene trap 1, H Earl Ruley|
|Allele Type||Gene trapped|
|Allele Synonym(s)||Pfdn1Gt(GTR1.3)1Rul; gene trap 1, H Earl Ruley|
|Gene Symbol and Name||Pfdn1, prefoldin 1|
|Gene Synonym(s)||expressed sequence AA408327; 2700086I23Rik; 2700086I23Rik; expressed sequence AU044714; PFD1; PDF; RIKEN cDNA 2700086I23 gene; AU044714; AA408327|
|Strain of Origin||129/Sv|
|Molecular Note||The retrovirus gene trap vector GTR1.3 containing a puromycin gene, a lacZ gene, a Pol2 gene and a neo cassette was inserted into intron 3. The absence of protein product was confirmed by western blot analysis on embryonic fibroblast extracts. Northern blot analysis confirmed the expression of a transcript of increased size in embryonic fibroblasts.|
When maintaining a live colony, heterozygous mice may be bred to wildtype mice from the colony or C57BL/6J inbred mice (Stock No. 000664). Homozygous mice die before five weeks of age due to neuromuscular dysfunction and physical wasting.
When using the Pfdn1- mouse strain in a publication, please cite the originating article(s) and include JAX stock #028765 in your Materials and Methods section.
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