Genetic Models of Calpain Deficiency and Ectopic Expression

Yan Gao; Christine Hall; James MacLeod; Peter A Greer

doi:10.1007/978-1-4939-8988-1_20

Genetic Models of Calpain Deficiency and Ectopic Expression

Methods Mol Biol. 2019:1915:261-274. doi: 10.1007/978-1-4939-8988-1_20.

Authors

Yan Gao^{1

2}, Christine Hall^{1

2}, James MacLeod^{1

2}, Peter A Greer^{3

4}

Affiliations

¹ Department of Pathology and Molecular Medicine, Queen's University, Kingston, ON, Canada.
² Division of Cancer Biology and Genetics, Cancer Research Institute, Kingston, ON, Canada.
³ Department of Pathology and Molecular Medicine, Queen's University, Kingston, ON, Canada. greerp@queensu.ca.
⁴ Division of Cancer Biology and Genetics, Cancer Research Institute, Kingston, ON, Canada. greerp@queensu.ca.

PMID: 30617810
DOI: 10.1007/978-1-4939-8988-1_20

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.

Keywords: CRISPR-Cas9 knockout; Calpain; Conditional Cre/loxP capns1 knockout mice; Lentiviral vectors; capn1; capn2; shRNA knockdown.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Calpain / deficiency
Calpain / genetics*
Cell Culture Techniques / methods*
Disease Models, Animal
Ectopic Gene Expression / genetics
Mice, Transgenic
Molecular Biology / methods*
Recombinant Proteins / genetics*

Substances

Recombinant Proteins
Calpain

Grants and funding

81189/CIHR/Canada