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

Evan A. Boyle,Johan O. L. Andreasson,Lauren M. Chircus,Samuel H. Sternberg,Michelle J. Wu,Chantal K. Guegler,Jennifer A. Doudna,William J. Greenleaf

Proceedings of the National Academy of Sciences（2016）

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摘要

The bacterial adaptive immune system CRISPR-Cas9 has been appropriated as a versatile tool for editing genomes, controlling gene expression, and visualizing genetic loci[1][1]. To analyze Cas9’s ability to bind DNA rapidly and specifically, we measured the kinetics of catalytically dead Cas9 (

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