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. 2023 Jun 29;150(12):dev201323. doi: 10.1242/dev.201323

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© 2023. Published by The Company of Biologists Ltd

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

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Fig. 1. — PCR tagging in zebrafish. (A) PCR tagging workflow: a gRNA/Cas9 RNP targeting the start or stop codon of a gene of interest is co-injected with a PCR-generated HDR template with short homology arms into fertilized zebrafish eggs, leading to integration of a fluorescent protein (FP) coding sequence. UTR, untranslated region. Red box indicates field of view in B,C and throughout the figures. (B,C) Lateral view of spinal cords of 3-5 dpf vamp2-mCherry and mEGFP-vamp1 PCR-tagged zebrafish showing vesicular fluorescence in synaptic boutons (arrowheads). Scale bars: 5 µm.