Abstract
Genetic manipulation in cell and animal models can provide important insights into gene function and the relationships between gene mutation and disease. This chapter describes methods to generate models of calpain-1 and calpain-2 deficiency, or their recombinant ectopic expression in cultured cells, and to genotype a conditional transgenic mouse model of calpain-1/calpain-2 deficiency that can be used to explore physiologic roles for these calpains.
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References
Wang H et al (2013) One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering. Cell 153(4):910–918
Tan Y et al (2006) Conditional disruption of ubiquitous calpains in the mouse. Genesis 44(6):297–303
Zufferey R et al (1998) Self-inactivating lentivirus vector for safe and efficient in vivo gene delivery. J Virol 72(12):9873–9880
Grieve S et al (2016) Calpain genetic disruption and HSP90 inhibition combine to attenuate mammary tumorigenesis. Mol Cell Biol 36(15):2078–2088
Shalem O et al (2014) Genome-scale CRISPR-Cas9 knockout screening in human cells. Science 343(6166):84–87
Sanjana NE, Shalem O, Zhang F (2014) Improved vectors and genome-wide libraries for CRISPR screening. Nat Methods 11(8):783–784
Acknowledgment
This work was supported by a grant from the Canadian Institutes of Health Research (81189) to PAG.
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Gao, Y., Hall, C., MacLeod, J., Greer, P.A. (2019). Genetic Models of Calpain Deficiency and Ectopic Expression. In: Messer, J. (eds) Calpain. Methods in Molecular Biology, vol 1915. Springer, New York, NY. https://doi.org/10.1007/978-1-4939-8988-1_20
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DOI: https://doi.org/10.1007/978-1-4939-8988-1_20
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