(American Association for the Advancement of Science, 2014-03-21) Lee, Je Hyuk; Daugharthy, Evan; Scheiman, Jonathan; Kalhor, Reza; Ferrante, Thomas; Yang, Joyce; Terry, Richard; Jeanty, Sauveur; Li, Chao; Amamoto, Ryoji; Peters, Derek; Turczyk, Brian; Marblestone, Adam; Inverso, Samuel; Bernard, Amy; Mali, Prashant; Rios, Xavier; Aach, John; Church, George
Understanding the spatial organization of gene expression with single nucleotide resolution requires localizing the sequences of expressed RNA transcripts within a cell in situ. Here we describe fluorescent in situ RNA sequencing (FISSEQ), in which stably cross-linked cDNA amplicons are sequenced within a biological sample. Using 30-base reads from 8,742 genes in situ, we examined RNA expression and localization in human primary fibroblasts using a simulated wound healing assay. FISSEQ is compatible with tissue sections and whole mount embryos, and reduces the limitations of optical resolution and noisy signals on single molecule detection. Our platform enables massively parallel detection of genetic elements, including gene transcripts and molecular barcodes, and can be used to investigate cellular phenotype, gene regulation, and environment in situ.
