A library of guide RNAs—each targeting a unique gene for CRISPR-based interference and carrying a unique barcode sequence—is introduced into a population of cells at a concentration that results in one guide RNA entering one cell, on average. Individual cells are then sorted into droplets bearing uniquely barcoded polyT primers, which are used to extract the cell’s mRNA. Sequencing the RNA then reveals both the introduced genetic mutations—determined by the guide RNA—and the transcriptional effect of that perturbation—determined by the collection of mRNAs bearing the cell-specific barcode (from the polyT primer).

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