C57BL/6J-Tg(Prnp-CHCHD2*)U105Jaaw/J

Stock number: 039978
Product name: D2^T61I
Strain synonyms: CHCHD2-T61I^U105
Research areas: Cell Biology Research, Metabolism Research, Neurobiology Research, Research Tools

Description

The D2^T61I transgenic mice express human coiled-coil-helix-coiled-coil-helix domain containing 2 (CHCHD2) with T61I mutation found in some Parkinson's disease (PD) patients, under the control of the murine Prnp (prion protein) promoter. These mice may be a useful mammalian model that recapitulates CHCHD2-T61I-mediated pathologies.

Detailed description

The D2^T61I transgenic mice harbor a transgene containing the human coiled-coil-helix-coiled-coil-helix domain (CHCHD2) with a T61I mutation, under the control of the murine Prnp (prion protein) promoter. The T61I mutations were found in the CHCHD2 gene in patients of Lewy body disorders (LBDs), including Parkinson's disease (PD). Hemizygous D2^T61I transgenic mice are viable and fertile, and they exhibit no gross abnormalities in brain and spinal cord morphology up to 12 months of age. Histological analysis at 12 months reveals reactive gliosis in the hippocampus and cortex, loss of synaptic integrity in the hippocampus, and mislocalization of α-synuclein. Hemizygous D2^T61I transgenic mice also exhibit motor deficits at 12 months of age without measurable cognitive impairment; however, by 18 months, cognitive deficits emerge, as assessed by novel object recognition testing. This line recapitulates key features associated with D2^T61I-linked pathology in human LBDs and may serve as a valuable model for mechanistic studies of disease-associated mutation.

Of note, in addition to these mice, the donating investigator provided The Jackson Laboratory Repository with D2^WT (Stock No. 039979).

Development

A D2^T61I transgene expression construct of ~14.8kb size consists of 3xFlag, human CHCHD2 cDNA (Reference sequence: NM_016139.4) cDNA sequence was first subcloned into the p3Xflag vector, which contains HindIII and SalI sites, in the same frame as the 3x flag tag. Then, the p.T61I (c.182 C>T) mutation was created by a pair of primers in the middle of which the T61I instead of the WT sequence was designed. After 2-step PCR and ligation, the T61I mutation was subcloned into the p3Xflag vector. Finally, the Flag-CHCHD2 sequences were subcloned into the MoPrp.xhoI vector (with an XhoI site added to Flag-CHCHD2-T61I) and the plasmid were sequenced for validity. The plasmid was linearized by Not1 and purified before microinjection. The construct was microinjected into at least 200 fertilized C57BL/6J embryos and implanted into pseudo-pregnant mice. Two potential founder lines, U144 and U105, were identified and bred with C57BL/6J mice for a more than five generations. They were then maintained on a C57BL/6J genetic background by the donating laboratory. This is the U105 line, and the integration site and copy numbers of the transgene are unknown. Upon arrival at The Jackson Laboratory, sperm were cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

Allele

Symbol: Tg(Prnp-CHCHD2*)U105Jaaw
Name: transgene insertion U105, Jung-A A Woo
MGI accession ID: MGI:7532877
Generation method: Transgenic
Attributes: Inserted expressed sequence; Epitope tag
Synonyms: CHCHD2-T61I^U105
Marker symbol: Tg(Prnp-CHCHD2*)U105Jaaw
Marker name: transgene insertion U105, Jung-A A Woo
Chromosome: UN
Strain of origin: C57BL/6J
Molecular note: The transgene insert contains the murine prion protein promoter and FLAG tag sequence in-frame with human CHCHD2 cDNA with a mutation that changes threonine codon 61(ACC) to isoleucine (ATC) (c.182C>T p.T61I). The mutation is found in some autosomal dominant Parkinson's disease (PD) patients.