Stumpf Craig R, Opperman Laura, Wickens Marvin
Program in Cellular and Molecular Biology, University of Wisconsin-Madison, Madison, Wisconsin, USA.
Methods Enzymol. 2008;449:295-315. doi: 10.1016/S0076-6879(08)02414-2.
RNA-protein interactions play an essential role in the maturation and regulation of RNAs within eukaryotic organisms. The three-hybrid system provides a simple, yet powerful means to study RNA-protein interactions within the eukaryote Saccharomyces cerevisiae. This chapter describes the basis of the system and applications in both examining specific RNA-protein interactions and screening libraries for novel interactions. We provide a detailed discussion on affinity versus reporter output, variations on library screening (e.g., randomization studies), some adaptations of the system, and updated reagents and protocols.
