How does it work?
ULTImate RNA Y3H is based on ULTImate Y2H, with modifications to use a small RNA as a bait. It consists of the detection of ‘bait-prey’ interactions thanks to the reconstitution of a full transcription factor from DNA Binding Domain and Activation Domain moieties. In the RNA Y3H approach, three hybrid molecules are used:
- A hybrid protein containing a DNA Binding Domain (DBD) fused to a RNA Binding Domain (RBD)
- A hybrid protein containing a transcriptional Activation Domain (AD) fused to a ‘prey’ protein fragment from the library
- A hybrid RNA molecule consisting of two RNA fragments, the RNA anchor interacting with RBD and the RNA bait.
Upon interaction of the RNA bait with the prey, the gap between the DBD and the AD is bridged, enabling the transcription of the reporter gene in yeast cells. The DNA of the positive clones is then sequenced and analyzed to identify the protein partners.
For more details about our screening technology and libraries, please refer to ULTImate Y2H and ULTImate Libraries.