Creative Biolabs is dedicated to providing yeast three-hybrid (Y3H) services including library construction and screening, which are introduced to assay RNA-protein interactions and devised to the selection of RNA binding protein with modifications to use a small RNA as bait. In 1996, the Wickens and Kuhl labs developed the yeast three-hybrid system independently. This system also makes it possible to identify those regions of an RNA or protein that are required for a known interaction and to test the combinations of RNA and protein to confirm whether they interact in vivo.
The Functions of Y3H
It is well-known that Y3H methods can be applied in a variety of research fields:
a. Finding protein partners of a known RNA sequence from cDNA libraries
b. Searching a natural RNA partner or ligand for a known RNA binding protein via RNA libraries
c. Testing suspected RNA-protein interactors
d. Mutational analysis of interacting RNA and protein
e. Discovery of multiprotein-RNA complexes
In addition, Y3H is still a sensitive method to screen for the interactions between small molecule drugs and their protein targets.
The Principle of Y3H
Yeast three-hybrid system is a derivative of yeast two-hybrid (Y2H). It’s a kind of powerful tool to dissert RNA-protein interactions of interest and that typically consists of three chimeric components.
The first hybrid protein is made up of an RNA binding protein (RBD) fused to a DNA binding domain (DBD). The second fusion protein molecule contains a second RNA binding protein fused to the transcriptional activation domain (AD). The third hybrid part is an RNA molecule which bridges above two fusion proteins by providing two specific RNA targets for the RNA binding proteins. When this tripartite constituent forms at a promoter, the reporter gene is turned on, even transiently. And the expressed reporter products can be recognized by simple biochemical or phenotypic assays.
Fig.1 General strategy of Y3H system.
Key Benefits of Y3H
In an RNA Y3H method, there are lots of great advantages presented on Creative Biolabs’ platforms:
RNA-protein interactions are essential for the proper execution and regulation of each step in the life of eukaryotic mRNAs, such as splicing, translational control, mRNA processing, and infection of RNA viruses. With this, Creative Biolabs provides a potentially effective technique by which to examine RNA-protein interactions towards a variety of purposes. The final submitted deliverables include an exhaustive report, a full list of identified protein partners, and interaction domain on each protein, together with scientific assistance whenever clients or their projects need for the better outcome.