FVB/N-Tg(Prnp-PFN1)34Zxu/J

Stock number: 028607
Product name: Prp-PFN1^WT transgenic line wt34 (or PrP-hPFN1wt line wt34)
Research areas: Cell Biology Research, Neurobiology Research, Research Tools

Description

Prp-PFN1^WT transgenic line wt34 (previously called PrP-hPFN1wt line wt34) has the mouse prion protein promoter directing expression of human wildtype profilin 1 (with N-terminal V5 tag) predominantly in both neurons and non-neuronal cells of the CNS. These mice are a control strain for Thy1.2-PFN1^C71G/Prp-PFN1^C71G transgenic mice (Stock No. 028608), and may be useful for studying the pathogenic role of PFN1 in familial amyotrophic lateral sclerosis (ALS or Lou Gehrig's disease).

Detailed description

Prp-PFN1^WT transgenic line wt34 has the mouse prion protein promoter directing expression of human wildtype profilin 1 with an N-terminal V5 tag (V5-PFN1^WT). This results in stable high expression of V5-PFN1^WT protein predominantly in the CNS (in both neurons and non-neuronal cells), with very low expression in the periphery. Hemizygous mice are viable and fertile with no reported neuromuscular abnormalities. The donating investigator reports they were not able to obtain homozygous mice to date (February 2016).

V5-PFN1^WT transgenic line wt34 is a control strain for the Thy1.2-PFN1^C71G/Prp-PFN1^C71G transgenic mice (Stock No. 028608); which has C71G mutant PFN1 expression leading to motor neuron degeneration and abnormalities in cytoskeleton, proteasome, autophagy and stress granules. These mice may be useful for studying the pathogenic role of PFN1 in familial amyotrophic lateral sclerosis (ALS or Lou Gehrig's disease).

Development

The Prp-PFN1^WT transgene (previously called PrP-hPFN1wt) was designed by Drs. Zuoshang Xu and John E. Landers (University of Massachusetts Medical School).

The PFN1^WT protein (V5-PFN1^WT) has a V5 tag (45 bp) fused to the N-terminus of a wildtype human profilin 1 cDNA sequence (PFN1; 423 bp / 141 aa). This V5-PFN1^WT sequence was inserted between exon 2 and exon 3 of mouse prion protein (PrP or Prnp) gene at a unique XhoI site in the MoPrP.Xho plasmid vector. The resulting 11,855 bp transgene was microinjected into FVB/NJ embryos. Transgenic founder male wt34 was bred with FVB/NJ females for germline transmission, and founder line wt34 was found to have stable high expression predominantly in the CNS. The integration site(s) and copy number of the Prp-PFN1^WT transgene is not known to date (February 2016). The Prp-PFN1^WT transgenic line wt34 colony was maintained by breeding hemizygotes with FVB/NJ mice for at least nine more generations, prior to sending one albino male mouse to The Jackson Laboratory Repository in 2016.

Upon arrival, sperm was cryopreserved. To generate our live colony, an aliquot of the frozen sperm was used to fertilize oocytes from FVB/NJ mice (Stock No. 001800). Thereafter, the colony was maintained by breeding hemizygous mice with wildtype (noncarrier) mice from the colony or to FVB/NJ mice (Stock No. 001800).

Allele

Symbol: Tg(Prnp-PFN1)34Zxu
Name: transgene insertion 34, Zuoshang Xu
MGI accession ID: MGI:5796265
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Marker symbol: Tg(Prnp-PFN1)34Zxu
Marker name: transgene insertion 34, Zuoshang Xu
Chromosome: UN
Strain of origin: FVB/NJ
Expressed genes: PFN1,profilin 1,human
Site of expression: Predominantly in the central nervous system.
Molecular note: The transgenic construct contains a human profilin 1 cDNA sequence with a V5 tag (45 bp) fused to the N-terminus and inserted between exon 2 and exon 3 of mouse prion protein gene at a unique XhoI site in the MoPrP.Xho plasmid vector. Founder line 34 was found to have stable high expression predominantly in the central nervous system.