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

**. 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- ...