The double ZZ changing the RNA world
RNAscope
The field of cancer immunotherapy has expanded rapidly in recent years with immune checkpoint inhibitors and other therapeutic approaches such as cancer vaccines and chimeric antigen receptor (CAR) therapy showing promising clinical results. Despite the dramatic and durable responses seen in many patients, our understanding of the immune response to cancer is still limited, and we cannot reliably predict who will or will not benefit from these new interventions. To better stratify patients for immunotherapy treatments, the series of events and biomarkers involved in the cancer‐immunity cycle need to be better understood. In addition, spatially mapped expression data at the single‐cell level is crucial to understanding the cellular organization and cell‐to‐cell interactions in the tumor and its complex microenvironment (TME).
RNAscope is a unique RNA ISH technology that provides single‐cell gene expression resolution with spatial and morphological context. The RNAscope assay detects mRNA and long non‐ coding RNAs in fresh-frozen, fresh fixed, and formalin‐fixed paraffin‐embedded (FFPE) cells and tissues by utilizing a unique double Z probe design and signal amplification strategy that allows for visualization of target RNA as a single dot, where each dot is an individual RNA molecule. The RNAscope strategy offers the key benefits of high sensitivity due to the signal amplification method, and high specificity because of the double Z probe design, resulting in a high signal‐to‐noise ratio in many tissues. The RNAscope assay enables single-molecule detection of vector DNA or transgene mRNA in the tissue at the single-cell level,
thus allowing for a high spatial resolution at both the cellular and subcellular level. Cell type-specific promoter activity and transgene expression heterogeneity can be assessed with the RNAscope assays, as can screening of vector delivery systems for cellular uptake efficiency and extracellular trapping of the vector.
