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Calpain pp 219–232Cite as

In Vivo Calpain Knockdown Using Delivery of siRNA

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1915))

Abstract

Calpain is an intracellular Ca2+-dependent non-lysosomal cysteine protease expressed ubiquitously in mammals. In endothelial cells, dysregulation of calpain has been shown to be involved in a wide variety of pathological conditions such as angiogenesis, vascular inflammation, and diabetes. Cell- or tissue-targeted in vivo delivery of small interfering RNA (siRNA) is a powerful research tool in the analysis of protein function and has been proposed as an attractive therapeutic modality that is applicable against a large number of human diseases including cancer. In this chapter we describe a method to knockdown calpain 1 in mouse pulmonary vascular endothelium using delivery of siRNA/cationic liposome complex. This technique results in a greater than 80% reduction in calpain 1 protein levels 48 h after a single i.v. injection of calpain 1 siRNA (0.5 mg siRNA/kg)/cationic liposome complex. We also describe confocal imaging to verify the loss of calpain 1 expression in pulmonary microvessel endothelial cells and application of this technique in the mouse model of ventilator-induced lung injury.

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Acknowledgments

This work was supported by the NIH Grant HL104092 to GH.

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Authors and Affiliations

  1. Department of Anesthesiology, University of Illinois College of Medicine, Chicago, IL, USA

    Xiaoqian Liu & Guochang Hu

  2. Department of Pharmacology, University of Illinois College of Medicine, Chicago, IL, USA

    Guochang Hu

Authors
  1. Xiaoqian Liu
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  2. Guochang Hu
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Corresponding author

Correspondence to Guochang Hu .

Editor information

Editors and Affiliations

  1. Department of Inflammation and Immunity, Lerner Research Institute, Cleveland Clinic, Cleveland, OH, USA

    Jeannette S. Messer

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Liu, X., Hu, G. (2019). In Vivo Calpain Knockdown Using Delivery of siRNA. 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_17

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  • DOI: https://doi.org/10.1007/978-1-4939-8988-1_17

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  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-4939-8987-4

  • Online ISBN: 978-1-4939-8988-1

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