Advancing Research in RNA-Based Therapeutics with Spatial Resolution
"The ability to directly visualize shRNA in brain tissue—with cellular precision—has advanced our therapeutic pipeline in ways no other technique could."
- Dr. Sonia Hesam-Shariati, PhD, School of Psychology, University of New South Wales
Resolving Spatial Distribution of shRNA in Complex Tissues
In our study of RNA-based therapeutics targeting long non-coding RNAs involved in addiction-related behaviors, one of the key challenges was confirming precise delivery and localization of shRNA within the brain. Traditional bulk RNA analysis methods, such as qPCR, could confirm expression but offered no insight into which cell types were targeted or where in the brain the therapy was active.
“Without RNAscope, it would be challenging to directly confirm these results and validate the success of our delivery system, making it an indispensable tool in our research.”
By applying an in-situ hybridization approach, we were able to detect shRNA expression at single-cell resolution, directly in brain, liver, and spleen tissues. This spatial resolution was critical for verifying that the therapeutic reached its intended cellular targets within highly heterogeneous tissue environments and helped refine our understanding of therapeutic distribution patterns.
Establishing Cellular Context for Therapeutic Efficacy
Behavioral outcomes in our models depend not just on whether RNA is expressed, but on its expression in the right cell types and brain regions. Using a method capable of detecting short RNA sequences with high specificity, we could evaluate the cell-type-specific uptake and localization of our shRNA following treatment.
This enabled us to link therapeutic presence to functional outcomes, distinguishing between on-target and off-target effects. Furthermore, combining this detection with other RNA markers allowed us to explore molecular interactions within defined anatomical regions, enhancing our interpretation of how RNA therapies influence behavior at the systems level.
“Being able to connect molecular localization with behavioral outcomes has significantly deepened our insight into how RNA-based therapies function in the brain.”