Current Approaches on Viral Infection: Proteomics and Functional Validations

Zheng, Jie; Tan, Boon Huan; Sugrue, Richard; Tang, Kai

Compared with siRNA knockdown, shRNA and miRNA mediated gene silencing were less reported as functional validation strategies to examine proteomic data. In one study, cellular c-Cbl was known to play a key role in macropinocytosis for entry of kaposi's sarcoma-associated herpesvirus (KSHV). Valiya Veettil et al. (2010) utilized MS to analyze the immunoprecipitation (IP) of c-Cbl and identified a novel interacting protein, myosin IIA. shRNA mediated knockdown of c-Cbl was then performed and the binding between c-Cbl and myosin IIA was abolished. Overall, these gene silencing studies showed that effective silencing of gene expression has a profound influence to functional virology studies in conjunction with proteomic techniques. What is more, it could provide a more rigorously demonstrated result at the functional level when siRNA-mediated knockdown is comprehensively combined with other approaches, such as immunofluorescence imaging, molecular cloning, RT-PCR, drug inhibition of secretory pathways.