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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. 1. — Characterization and activity of CRISPR-ErCas12a in zebrafish. (A) Phylogenetic relationship between known CRISPR associated proteins and ErCas12a. Evaluation of branching was ensured by bootstrap statistical analysis (1,000 replications). (B) Workflow showing dual NLS ErCas12a mRNA and pre-crRNA injection into single-cell animals. Animals are heat shocked for 4 h and then allowed to develop normally until DNA is isolated and analyzed. (C) Schematic and sequences of pre-crRNA used to target noto. (D) Schematic and sequences of pre-crRNA used to target cx43.4. (E) Results displaying the wild-type and top five mutated alleles after AmpliconEZ analysis of two noto pre-crRNAs and one cx43.4 pre-crRNA.