Advertisement

Calpain pp 261-274 | Cite as

Genetic Models of Calpain Deficiency and Ectopic Expression

  • Yan Gao
  • Christine Hall
  • James MacLeod
  • Peter A. GreerEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1915)

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.

Key words

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

Notes

Acknowledgment

This work was supported by a grant from the Canadian Institutes of Health Research (81189) to PAG.

References

  1. 1.
    Wang H et al (2013) One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering. Cell 153(4):910–918CrossRefGoogle Scholar
  2. 2.
    Tan Y et al (2006) Conditional disruption of ubiquitous calpains in the mouse. Genesis 44(6):297–303CrossRefGoogle Scholar
  3. 3.
    Zufferey R et al (1998) Self-inactivating lentivirus vector for safe and efficient in vivo gene delivery. J Virol 72(12):9873–9880PubMedPubMedCentralGoogle Scholar
  4. 4.
    Grieve S et al (2016) Calpain genetic disruption and HSP90 inhibition combine to attenuate mammary tumorigenesis. Mol Cell Biol 36(15):2078–2088CrossRefGoogle Scholar
  5. 5.
    Shalem O et al (2014) Genome-scale CRISPR-Cas9 knockout screening in human cells. Science 343(6166):84–87CrossRefGoogle Scholar
  6. 6.
    Sanjana NE, Shalem O, Zhang F (2014) Improved vectors and genome-wide libraries for CRISPR screening. Nat Methods 11(8):783–784CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Yan Gao
    • 1
    • 2
  • Christine Hall
    • 1
    • 2
  • James MacLeod
    • 1
    • 2
  • Peter A. Greer
    • 1
    • 2
    Email author
  1. 1.Department of Pathology and Molecular MedicineQueen’s UniversityKingstonCanada
  2. 2.Division of Cancer Biology and GeneticsCancer Research InstituteKingstonCanada

Personalised recommendations