C57BL/6J-Tg(Prnp-CHCHD2)U27Jaaw/J

Stock number: 039979
Product name: D2^WT
Strain synonyms: CHCHD2-WT
Research areas: Cell Biology Research, Metabolism Research, Neurobiology Research, Research Tools

Description

The D2^WT transgenic mice express wild type human coiled-coil-helix-coiled-coil-helix domain containing 2 (CHCHD2) under the control of the murine Prnp (prion protein) promoter. These mice may be useful for studying the CHCHD2-mediated pathologies found in some Parkinson's disease (PD) patients.

 

Detailed description

The D2^WT transgenic mice express the human CHCHD2 gene under the control of the murine Prnp (prion protein) promoter. Mutations in CHCHD2 have been implicated in patients with Lewy body disorders (LBDs), including Parkinson's disease (PD). Hemizygous D2^WT mice are viable and fertile, with no gross abnormalities in brain and spinal cord morphology observed up to 12 months of age. By 12-months, there is approximately a 2.3-fold increase in total Chchd2 transcripts (mouse and human), with widespread transgene expression in the cortex, midbrain, hippocampus, and spinal cord. Unlike D2^T61I mice, D2^WT mice do not show cognitive deficits at 18 months of age. Furthermore, aged D2^WT mice exhibit relative protection from age-associated neuroinflammation, insoluble protein aggregation, and ultrastructural abnormalities of mitochondria and lysosomes, when compared to C57BL/6J controls. This line may serve as a model of CHCHD2-WT overexpression to investigate its role in neuroprotection and age-related brain homeostasis, and as a comparison control for studies using the D2^T61I disease model.

Of note, in addition to these mice, the donating investigator provided The Jackson Laboratory Repository with D2^T61I (Stock No. 039978).

Development

A D2^WT transgene expression construct of ~14.8kb size consists of 3xFlag, human CHCHD2 cDNA, subcloned into the XhoI site of the MoPrp. Specifically, the wild type human CHCHD2 cDNA sequence (Reference sequence: NM_016139.4) was first subcloned into the p3Xflag vector, which contains HindIII and SalI sites, in the same frame as the 3x flag tag. Finally, the Flag-CHCHD2 sequences were subcloned into the MoPrp.xhoI vector (with an XhoI site added to Flag-CHCHD2-WT) 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. One positive founder line, U27, was identified and bred with C57BL/6J mice for more than five generations. The line was maintained on a C57BL/6J genetic background by the donating laboratory. 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)U27Jaaw
Name: transgene insertion U27, Jung A Woo
MGI accession ID: MGI:8226206
Generation method: Transgenic
Attributes: Inserted expressed sequence; Epitope tag
Synonyms: D2^WT
Marker symbol: Tg(Prnp-CHCHD2)U27Jaaw
Marker name: transgene insertion U27, Jung 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.