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. 2019 Dec 16;2(6):417–433. doi: 10.1089/crispr.2019.0026

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© Wesley A. Wierson et al. 2019; Published by Mary Ann Liebert, Inc.

This Open Access article is distributed under the terms of the Creative Commons License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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FIG. 2. — Using noto:RFP-DR48 to assay the propensity of ErCas12a and SpCas9 to elicit strand annealing in zebrafish. (A) Schematic of noto:RFP-DR48 showing the location of the 48 bp direct repeats flanking both the SpCas9 and ErCas12a universal RNA cursor sites (underline) and protospacer adjacent motif (PAM) sites (red text). (B) Data plot showing the ratio of injected animals displaying red fluorescent protein (RFP) in the notochord out of the total carrying the transgene. Data plot represents the mean ± SD. p-Values calculated with one-tailed Student's t-test. (C) Qualitatively scored ratios of notochord converted to RFP+ after SpCas9 induced strand-annealing mediated repair (SAMR). (D) Qualitatively scored ratios of notochord converted to RFP+ after ErCas12a induced SAMR. (E–G and E'-G') Representative embryos for broad, intermediate, and narrow conversion of RFP in the notochord.