Screening for Host Factors Directly Interacting with RSV Protein: Microfluidics

Methods Mol Biol. 2016:1442:165-74. doi: 10.1007/978-1-4939-3687-8_12.

Abstract

We present a high-throughput microfluidics platform to identify novel host cell binding partners of respiratory syncytial virus (RSV) matrix (M) protein. The device consists of thousands of reaction chambers controlled by micro-mechanical valves. The microfluidic device is mated to a microarray-printed custom-made gene library. These genes are then transcribed and translated on-chip, resulting in a protein array ready for binding to RSV M protein.Even small viral proteome, such as that of RSV, presents a challenge due to the fact that viral proteins are usually multifunctional and thus their interaction with the host is complex. Protein microarrays technology allows the interrogation of protein-protein interactions, which could possibly overcome obstacles by using conventional high throughput methods. Using microfluidics platform we have identified new host interactors of M involved in various cellular pathways. A number of microfluidics based assays have already provided novel insights into the virus-host interactome, and the results have important implications for future antiviral strategies aimed at targets of viral protein interactions with the host.

Keywords: Integrated microfluidics; Protein arrays; RSV–host interaction; Virus–host interactions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Gene Library
  • HEK293 Cells
  • Host-Pathogen Interactions
  • Humans
  • Microfluidic Analytical Techniques / instrumentation*
  • Printing, Three-Dimensional
  • Protein Array Analysis / instrumentation
  • Protein Array Analysis / methods
  • Protein Interaction Mapping / instrumentation
  • Protein Interaction Mapping / methods*
  • Respiratory Syncytial Virus Infections / metabolism*
  • Respiratory Syncytial Virus Infections / virology
  • Respiratory Syncytial Virus, Human / metabolism
  • Respiratory Syncytial Virus, Human / physiology*
  • Viral Matrix Proteins / metabolism*

Substances

  • Viral Matrix Proteins