Technologies
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WARF: P04156US

Assay to Detect Viral Uncoating


INVENTORS -

John Young, Shakti Narayan

The Wisconsin Alumni Research Foundation (WARF) is seeking commercial partners interested in developing a cell-free system for studying viral uncoating.
OVERVIEWDuring or shortly after a retrovirus particle enters a host cell, the particle undergoes a process known as “uncoating,” where the viral genome is freed from the viral envelope and becomes accessible for reverse transcription. Because uncoating is required for viral infection and replication, understanding how it works is of major interest. However, a key hurdle in the study of uncoating is the lack of a cell-free system that reproduces retroviral fusion, uncoating, and reverse transcription in vitro.
THE INVENTIONUW-Madison researchers have developed a cell-free system for studying the viral uncoating mechanism. This system uses a sub group of avian sarcoma and leucosis virus (ASLV-A) as a model retrovirus. The TVA receptor for ASLV-A is incorporated into the human cell line 293. After ASLV-A enters the 293 cells and accumulates in the cell’s endosomes, the cells are lysed. The virus-containing endosomes are isolated under conditions of high pH, which block entry of the virus into the cytosol, providing a cell-free system for studying uncoating. Conditions can then be altered to determine the effect of various agents on viral uncoating and other post viral envelope-endosome fusion events. Experiments have shown that the viral nucleic acid released from the endosomes is competent for reverse transcription, and that viral DNA synthesis is dependent on cellular factors contained in the S100 or S10 fraction of the 293 cell line.
APPLICATIONS
  • Screening anti-viral compounds that block retroviral uncoating
  • Retroviral uncoating studies
  • May lead to the discovery of cellular factors or other compounds that play a role in uncoating
  • May be used to screen for HIV inhibitors
KEY BENEFITS
  • Provides the first cell-free viral uncoating system
  • Applicable to many retroviral systems, including HIV, SIV, HTLV, MPMV, MLV, and ALV
  • Amenable to high throughput screening
ADDITIONAL INFORMATION
Contact Information
For current licensing status, please contact Rafael Diaz at rdiaz@warf.org or 608-960-9847.
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