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Researchers at Stanford have developed the SNAIL-RCA method for inexpensive and efficient multiplexed detection of single RNA molecules in single cells.
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Background: Researchers at Stanford have developed the SNAIL-RCA method for inexpensive and efficient multiplexed detection of single RNA molecules in single cells. Current methods to analyze the abundance and spatial distribution of mRNA are inefficient, expensive, and time consuming. New methods are needed. To help meet this need the inventors developed the SNAIL-RCA (Splint Nucleotide Assisted Intramolecular Ligation followed by Rolling Circle Amplification) method. It provides a simplified proximity ligation technique to allow multiplexed, quantitative, targeted detection of RNA molecules with single-cell resolution. The RCA products can be detected with a variety of techniques including optical microscopy and SIMS ion beam imaging. This technology will aide biomarker discovery and enable development of molecular diagnostics, including cancer diagnostics, for precision medicine. Stage of research: Validation studies are ongoing and show great promise. Albumin mRNA in mouse liver tissue section Applications: Multiplexed and quantitative gene expression profiling in single cells for: Molecular diagnostics for precision medicine Biomarker discovery Research Advantages: Allows simultaneous analysis of multiple nucleic acids and proteins in a single cell Simple design of SNAIL-RCA: Requires only a handful of probe pairs per gene Allows for simple configuration of oligonucleotides Eliminates need for boilerplate hybridization-ligation sequences Reduces the overall reagent cost per gene Streamlined proximity ligation protocol Efficient detection of small RNAs Method is compatible with conventional antibody staining Opens the possibility for much higher degree of multiplexing