WARF TECHNOLOGIES

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.

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

• 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.

For More Information About the Inventors

• Marvin Wickens

Tech Fields

• Research Tools : Protein interactions & function