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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. 3. — Targeting noto with GeneWeld. (A) Schematic of noto showing designed homology for precise integration using ErCas12a and the U-pre-crRNA. Green is designed 5′ homology. Blue is designed 3′ homology. The PAM for ErCas12a targeting in the genome and donor is underlined. (B and B′) Confocal Z-stack image showing broad green fluorescent protein (GFP) expression in the embryo. Scale bar is 100 μm. (C) Data plot showing the ratio of embryos with GFP expression in the notochord out of total injected embryos. Data plot represents the mean ± SD. (D) Gel showing 5′ junction fragment expected after precise integration using GeneWeld. (E) Gel showing 3′ junction fragment expected after precise integration using GeneWeld. (F) DNA sequencing of lane 5 in (D) and (E) showing a precise integration using the programmed homology.