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bioRxiv · 10.1101/059782

High-throughput biochemical profiling reveals Cas9 off-target binding and unbinding heterogeneity

Evan August Boyle·Johan Oscar Lennart Andreasson·Lauren Mallory Chircus·Samuel Henry Sternberg·Michelle Jennifer Wu·Chantal Katrin Guegler·Jennifer Anne Doudna·William James Greenleaf
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Abstract

**. Introduction **. Introduction **. Main body **. Online Methods **. Author contributions **. Competing financial... ** References The bacterial adaptive immune system CRISPR-Cas9 has been appropriated as a versatile tool for editing genomes, controlling gene expression, and visualizing genetic loci1. To analyze Cas9s ability to bind DNA rapidly and specifically, we measured the kinetics of catalytically dead Cas9 (dCas9) interactions with a library of potential binding partners. Using a massively parallel assay of protein-DNA interactions derived from a high-throughput sequencing flow cell (HiTS-FLIP)2 and building on the established importance of protospacer adjacent motif (PAM) and seed recognition3- ...

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Evan August Boyle, Johan Oscar Lennart Andreasson, Lauren Mallory Chircus, Samuel Henry Sternberg, Michelle Jennifer Wu, Chantal Katrin Guegler, Jennifer Anne Doudna, William James Greenleaf. 2016-06-19. High-throughput biochemical profiling reveals Cas9 off-target binding and unbinding heterogeneity. https://doi.org/10.1101/059782

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