Wisconsin Alumni Research Foundation

Research Tools
Research Tools
Strain and Vector for Use with the Three Hybrid System
WARF: P97099US

Inventors: Marvin Wickens, Beilin Zhang

The Wisconsin Alumni Research Foundation (WARF) is seeking commercial partners interested in developing an improved three hybrid system.
Three hybrid screens are used to detect RNA-protein interactions.  A problem with three hybrid screens has been high background due to some cDNAs binding and activating transcription in the absence of a RNA-protein interaction.
The Invention
UW-Madison researchers have corrected this problem by the addition of the screening marker ade2 onto the plasmids containing the hybrid RNA and the ura3 marker.  They created a yeast cell line containing the sequence Lex A-CP-CP (where CP is coat protein) in its genome, and plasmids that encode a hybrid RNA molecule and ura3 as a selectable marker.

The researchers have utilized this three hybrid system to detect RNA-protein interactions as measured by reporter function activity, such as transcription of a his3 reporter gene. The most common application is one in which the RNA is known and interacting proteins unknown. In this case, a library of cDNAs is screened to detect a protein of interest.
  • Three hybrid screens
Key Benefits
  • This new strain and plasmid construct provide a means for overcoming the high background/DNA binding problems associated with the three hybrid system.
  • Superfluous portions of the RNA-binding CPs that are not involved in RNA binding have been removed, further contributing to the decrease in background.
Additional Information
For More Information About the Inventors
For current licensing status, please contact Jennifer Gottwald at [javascript protected email address] or 608-960-9854