Summary: RNAscope™ in situ hybridization (ISH) is widely used to detect mRNA in tissue sections and is often paired with a protein marker. The RNAscope™ Plus assay enables detection of one siRNA/ASO, up to three mRNAs, and one protein for cell-specific analysis of biodistribution and target knock-down—within the same tissue section. When integrated with the PhenoImager HT system (Akoya Biosciences), autofluorescence can be leveraged to train AI classifiers for advanced image analysis. A new protease-free RNAscope workflow now allows detection of 1 siRNA, 1–3 mRNAs, and 2–4 proteins via sequential protein detection using the TSA workflow (Akoya), creating a powerful multiomic tissue analysis solution for oligonucleotide programs.
Learning Objectives
- See practical experience using RNAscope Plus for detection of oligonucleotides (siRNA/ASO) in tissue.
- Set up 6-plex RNAscope Plus assays for simultaneous detection of siRNA, mRNAs, and proteins across tissues.
- Analyze RNAscope Plus data with Visiopharm to classify tissue, quantify siRNA, and measure target knock-down.
Speaker
Louis Szeponik, Ph.D.
Senior Scientist, Discovery Imaging Sciences, AstraZeneca (Sweden)
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RNAscope™ and ACD are trademarks of Advanced Cell Diagnostics (ACD), a Bio-Techne brand. PhenoImager™ and TSA™ are trademarks of Akoya Biosciences. Visiopharm® is a trademark of Visiopharm. Company and product names are the property of their respective owners.