Investigation of the functional roles of host cell proteins involved in coronavirus infection using highly specific and scalable RNA interference (RNAi) approach.

Abstract

Since its identification in the 1990s, the RNA interference (RNAi) pathway has proven extremely useful in elucidating the function of proteins in the context of cells and even whole organisms. In particular, this sequence-specific and powerful loss-of-function approach has greatly simplified the study of the role of host cell factors implicated in the life cycle of viruses. Here, we detail the RNAi method we have developed and used to specifically knock down the expression of ezrin, an actin binding protein that was identified by yeast two-hybrid screening to interact with the Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) spike (S) protein. This method was used to study the role of ezrin, specifically during the entry stage of SARS-CoV infection.

• Authors: Jean Kaoru Millet, Béatrice Nal
• Journal: Methods in molecular biology (Clifton, N.J.) (Methods Mol Biol) Vol. 1282 Pg. 231-40 ( 2015) ISSN: 1940-6029 [Electronic] United States
• PMID: 25720484 (Publication Type: Journal Article)

Chemical References

• Cytoskeletal Proteins
• ezrin

Topics

• Cytoskeletal Proteins (physiology)
• Gene Knockdown Techniques
• HeLa Cells
• Host-Pathogen Interactions
• Humans
• RNA Interference
• Severe acute respiratory syndrome-related coronavirus (physiology)
• Virus Internalization