B6SJL-Tg(Prnp-Immt/SOD1)1Gmnf/J

Stock number: 025402
Product name: mito-WTSOD1
Research areas: Cell Biology Research, Neurobiology Research, Research Tools

Description

mito-WTSOD1 transgenic mice express mitochondrial inner membrane-tethered, human wildtype SOD1 at high levels in brain, spinal cord, heart and skeletal muscle. These mito-WTSOD1 mice are a control strain for mito-G93ASOD1 mice (Stock No. 024503), and may be useful for studying the pathogenic role of SOD1 and mitochondria in familial amyotrophic lateral sclerosis (ALS or Lou Gehrig's disease).

Detailed description

mito-WTSOD1 transgenic mice have the mouse prion protein promoter directing expression of wildtype human SOD1 with N-terminal fusion of the mouse mitofilin mitochondrial targeting signal, MMP cleavage site and transmembrane domain. This results in expression of SOD1 that is anchored to the outer side of the mitochondrial inner membrane (i.e., facing the intermembrane space) at high levels in brain, spinal cord, heart and skeletal muscle. High expression is observed in both neurons and astrocytes. Low to undetectable levels of protein are reported in kidney, lung, spleen and liver. The mito-SOD1 protein in high-expressing tissues oligomerizes and acquires enzymatic activity. Hemizygous mito-WTSOD1 mice are viable and fertile with no reported neuromuscular abnormalities. The donating investigator reports that homozygotes were not obtained and may not be viable.

These mito-WTSOD1 mice are a control strain for mito-G93ASOD1 mice (Stock No. 024503). In addition, when mito-WTSOD1 mice are bred to Sod1-deficient mice (such as Stock No. 002972), the resulting mitoSOD1,Sod1^-/- animals are protected from the motor axonopathy and mitochondrial abnormalities observed in Sod1^-/- motor neurons.

Development

The PrP-mitoSOD1^WT transgene (PrP-mito-WTSOD1 or mitoSOD1) was designed by Drs. Giovanni Manfredi and Jordi Magrane (Weill Medical College of Cornell University). The mito-WTSOD1 fusion protein contains a mouse mitofilin (Immt) cDNA sequence encoding the first 187 amino acids (including the mitochondrial targeting presequence, MMP cleavage site and transmembrane domain) fused in-frame to the N-terminus of a wildtype human Cu,Zn superoxide dismutase (SOD1) cDNA sequence. The SOD1 start codon was removed and substituted with an in-frame 6 nt DNA linker. This mito-WTSOD1 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. In contrast to the publication, the donating investigator reports the resulting PrP-mito-WTSOD1 transgene was microinjected into [C57BL/6J x CBA/J]F2 fertilized eggs. Male transgenic founders were bred with B6SJLF1/J female mice, and and the founder line with the highest expression level (founder line 1) was characterized. At that time, quantitative real-time PCR determined transgenic animals had ~45 transgene copies, and breeding performance indicated autosomal integration of the transgenes. The mito-WTSOD1 transgenic colony was maintained by breeding hemizygous males with B6SJLF1/J females for at least ten generations prior to sending males to The Jackson Laboratory Repository in 2014. Upon arrival, sperm was cryopreserved. To generate our live colony, an aliquot of the frozen sperm was used to fertilize oocytes from B6SJLF1/J mice (Stock No. 100012). Thereafter, the colony was maintained by breeding hemizygous mice with B6SJLF1/J mice every generation.

Allele

Symbol: Tg(Prnp-Immt/SOD1)1Gmnf
Name: transgene insertion 1, Giovanni Manfredi
MGI accession ID: MGI:5575877
Generation method: Transgenic
Attributes: Inserted expressed sequence; Humanized sequence
Marker symbol: Tg(Prnp-Immt/SOD1)1Gmnf
Marker name: transgene insertion 1, Giovanni Manfredi
Chromosome: UN
Strain of origin: (C57BL/6 x CBA)F1
Expressed genes: Immt,inner membrane protein, mitochondrial,mouse, laboratory; SOD1,superoxide dismutase 1,human
Molecular note: The mito-WTSOD1 fusion protein contains a mouse mitofilin (Immt) cDNA sequence encoding the first 187 amino acids (including the mitochondrial targeting presequence, MMP cleavage site and transmembrane domain) fused in-frame to the N-terminus of a wild-type human Cu,Zn superoxide dismutase (SOD1) cDNA sequence. The SOD1 start codon was removed and substituted with an in-frame 6 nt DNA linker. This mito-WTSOD1 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 founder line with the highest expression level (founder line 1) was characterized.