Health
Inhibition of CRISPR-Cas12a DNA targeting by nucleosomes and chromatin – Science Advances
Genome engineering nucleases must access chromatinized DNA. Here, we investigate how AsCas12a cleaves DNA within human nucleosomes and phase-condensed nucleosome…
INTRODUCTION
Class 2 CRISPR-Cas nucleases Cas12a and Cas9 are precision genome-editing tools that are evolutionarily and structurally distinct yet share several key reaction steps (13). Class 2 nucleases first identify a protospacer adjacent motif (PAM) via protein-DNA contacts. PAM binding causes local DNA unwinding and initiates directional strand invasion by the CRISPR RNA (crRNA) (4). An R-loop between the crRNA guide sequence and the complementary target DNA extends toward the PAM-distal end…
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