Assessing differential expression of protein interactions and RNA by combining isPLA and HCR™ Pro RNA-FISH on the BOND RX automated research stainer

The advent of the spatial omics era has enabled researchers to acquire vast amounts of transcriptomics and proteomics data. The abundance of a transcript, however, does not necessarily display strong correlation with protein expression levels, so a holistic approach to sample analysis is crucial.

 

HCR™ Pro RNA-FISH by Molecular Instruments enables high-resolution, multiplex detection of transcripts without requiring harsh protease-digestion steps. The in situ proximity ligation assay technology from Navinci allows the study of protein functional states by sensitively detecting protein-protein interactions. Both technologies can be fully and robustly automated on the BOND RX automated research stainer by Leica Biosystems.

 

For the first time, we have combined two state-of-the art technologies for detecting RNA expression and protein interactions to better understand tissue phenotypes from a functional perspective. As a proof of principle, we focused on the PD1/PD-L1 inhibitory pathway, detecting the PD1 and PD-L1 transcripts, as well as the interaction between them on a protein level.

 

Learning Objectives