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. 2021 Feb 22;49(5):2973–2984. doi: 10.1093/nar/gkab105

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© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 4. — Mechanism of prespacer splay and length determination. (A) D10 functions as a wedge to terminate the duplex region of the prespacer. The Grabber motif of Q72 and F73 bends and guides the single stranded 3′-overhang to the active site of Cas1. (B) Close up of the splayed prepsapcer showing the wedge residue D10 and the Grabber motif. (C) The 26-bp duplex of 26–5–5 prespacer is unwounded by residue D10 of Cas1, leading to a 22-bp duplex. (D) The displaced 5′ overhang of prepsapcer interacts with the N-terminal domain of Cas1.