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. Author manuscript; available in PMC: 2020 Jan 8.
Published in final edited form as: Nat Biotechnol. 2018 Nov 27:10.1038/nbt.4317. doi: 10.1038/nbt.4317

Figure 1. Mutational profiles generated by CRISPR/Cas9, and a method for their high-throughput measurement.

Figure 1

High throughput measurement of repair outcomes. Constructs containing both a gRNA and its target sequence (matched colors) in variable context (grey boxes) are cloned en masse into target vectors containing a human U6 promoter (green) (1), packaged into lentiviral particles, and used to infect cells (2), where they generate mutations at the target (3). DNA from the cells is extracted, the target sequence in its context amplified with common primers, and the repair outcomes (location, size, and sequence of mutation) determined by deep short read sequencing (4).