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DNA interrogation by the CRISPR RNA-guided endonuclease Cas9

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DNA interrogation by the CRISPR RNA-guided
endonuclease Cas9
Samuel H. Sternberg, Sy Redding, Martin Jinek, Eric C. Greene &
Jennifer A. Doudna
2014

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Martin Jinek, Jennifer A. Doudna
Berkeley, California

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CRISPR/Cas9 bacterial immune system

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1. Single-molecule visualization of Cas9
NIHMS555417-supplement-12.mov

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1. Single-molecule visualization of Cas9

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2. Cas9-RNA finds targets by 3D diffusion
NIHMS555417-supplement-13.m4v

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3. A PAM is required for DNA interrogation

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4. Mechanism of RNA-DNA heteroduplex formation

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5. The PAM triggers Cas9 nuclease activity
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