Unmet Need Traditional gene expression methods that use bulk RNA analysis lack the ability to resolve transcript location, and thus fail to capture the inherent cellular heterogeneity present in virtually every tissue. Abundance and localization of RNA transcripts mapped to specific regions of tissue can complement histological analysis, providing an additional layer of molecular information. Subcellular resolution of mRNA abundance and location can in some cases be used to categorize biologically important cell-to-cell variability and interactions. Technology Overview The field of in situ spatial biology is revolutionizing our understanding of tissue architecture, disease processes and immune responses. The LISH technology described above has been further developed into LISH-Lock'n'Roll (LISH-LnR), a multiplexed probe ligation technology that uses bipartite probe sets and rolling circle amplification for RNA detection in situ, which enables detection and discrimination of even highly similar RNA sequences with subcellular resolution. LISH-LnR allows for cost-effective, simultaneous detection of a large number of RNA sequences with improved specificity and sensitivity, overcoming many limitations of existing methods. Additionally, Dr. Larman has developed a system of “molecular rheostats” that tune the LISH-LnR assay performance, a complementary sequencing-based assay for probe panel qualification (using LISH-seq C13639), and a QC method to determine the quality and availability of fixed tissue RNA (using LISH-QC C13639). The LISH-LnR work is described in a new publication. The associated patent application can be found here. A patent application covering an improved version of the LISH-LnR technology has also been filed here.
Stage of Development Efficacy and specificity of “Lock’n’Roll” target multiplexing has been tested. In vitro data is available.
