Basic Information
Description
The Pink1 gene encodes a mitochondrially located serine/threonine kinase. Analysis in Pink1 deficient mice suggests Pink1 is important for mitochondrial function and provides critical protection against both intrinsic and environmental stress, suggesting a pathogenic mechanism by which loss of Pink1 may lead to nigrostriatal degeneration in PD (Parkinson’s disease).
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Targeting Strategy
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Pink1 knockout rats were developed on a Sprague-Dawley background. Two sgRNAs were designed to generate a ~ 6.5-kb deletion at the site of exons 2 – 4 in the rat genome such that most of the coding region of the Pink1 gene was knocked out. Offspring were maintained on a pure Sprague-Dawley strain genetic background by breeding homozygous males and females
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Details
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Genotyping Information
Primer Sequence (5’-3’) Tm (℃)
Product size (bp) Pink1KO primer 1 TACTGACTTCTCGACCCAACTT 62 WT: 7109 Mut: ~400
Pink1KO primer 2 GAACTTGTGGCCGTTTACGTCG 62 Polymerase: KOD-plus
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References
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- Gandhi, S., Wood-Kaczmar, A., Yao, Z., Plun-Favreau, H., Deas, E., Klupsch, K., Downward, J., Latchman, D. S., Tabrizi, S. J., Wood, N. W., Duchen, M. R., Abramov, A. Y. 2009. PINK1-associated Parkinson’s disease is caused by neuronal vulnerability to calcium-induced cell death. Molec. Cell 33: 627-638. [PubMed: 19285945]
- Tang, B., Xiong, H., Sun, P., Zhang, Y., Wang, D., Hu, Z., Zhu, Z., Ma, H., Pan, Q., Xia, J., Xia, K., Zhang, Z. 2006. Association of PINK1 and DJ-1 confers digenic inheritance of early-onset Parkinson’s disease. Hum. Molec. Genet. 15: 1816-1825. [PubMed: 16632486]
- Valente, E. M., Abou-Sleiman, P. M., Caputo, V., Muqit, M. M. K., Harvey, K., Gispert, S., Ali, Z., Del Turco, D., Bentivoglio, A. R., Healy, D. G., Albanese, A., Nussbaum, R., and 10 others. 2004. Hereditary early-onset Parkinson’s disease caused by mutations in PINK1. Science 304: 1158-1160. [PubMed: 15087508]
- Yang, Y., Gehrke, S., Imai, Y., Huang, Z., Ouyang, Y., Wang, J.-W., Yang, L., Beal, M. F., Vogel, H., Lu, B. 2006. Mitochondrial pathology and muscle and dopaminergic neuron degeneration caused by inactivation of Drosophila Pink1 is rescued by parkin. Proc. Nat. Acad. Sci. 103: 10793-10798. [PubMed: 16818890]
