Rainbow UltraFISH: Rapid Sequential RNA FISH for Single-cell Gene Expression Analysis

SBIR AbstractSingle cell biology is a multi disciplinary study to investigate how large numbers of molecular species interact with each other in single cellsand how large population of cells work together as complex systemsCells within a given tissueeven when they are morphologically similarare heterogeneous in their gene expression patternsDefining gene expression patterns at the single cell level within the context of the normal tissue architecture is essential to understanding tissue function and developmentand will ultimately benefit precision and personalized medicineHowevercurrent tools for single cell gene expression analysissuch as single cell RNA sequencinglack spatial information and have low cell and RNA detection efficiencyHencethere is an urgent need for better tools for profiling single cell gene expression patterns in situi ewithin the context of normal tissue architecture and cell cell relationshipsRainbow UltraFISH is a technology that integrates spatial and sequence information together for single cell RNA analysisIt uses rapid sequential fluorescent in situ hybridizationFISHto achieve multiplex RNA detection at high detection efficiency and singlecell resolutionIn cultured cell linesUltraFISH has been shown to complete each round of probe target hybridization in andltminwith a detection efficiency ofIn this proposalwe will expand the UltraFISH technology into primary mouse tissue samplesincluding fresh frozen tissueAimformaldehyde fixedfrozen tissueAimand formalin fixedparaffin embedded tissueAimWith the successful outcome of this proposalUltraFISH will be the simplest and fastest technology for single cell gene expression analysis in situ and ultimately enable more precision medicine SBIR Project Narrative Project NarrativeRainbow UltraFISH is a technology that will enable researchers to analyze the expression patterns of hundreds to thousands of genes within single cells in the context of their normal tissue environmentWith the coming of the spatial genomics eraUltraFISH will become an essential research tool for a very broad range of biologists and medical scientists in both academia and the biotech industryIt will help answer fundamental questions in all aspects of biologyas well as serve as a powerful tool to unravel the secrets of human disease