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. 2019 Aug 8;47(16):8632–8648. doi: 10.1093/nar/gkz677

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© The Author(s) 2019. 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 License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Cas1–Cas2 spacer integration reaction is directional. (A) Detection of full-site and half-site integration products with spacer hairpin CRISPR targets (top panel) and leader hairpin CRISPR targets (bottom panel) with unmodified (OH/OH) and modified (OH/dd, dd/OH, dd/dd) pre-spacers show site of the first transesterification reaction. (B) Time-course of spacer integration assay using unmodified pre-spacer and spacer hairpin CRISPR target. Quantification of B (bottom panel).