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

Fig. 4.

Fig. 4.

N-terminal Vamp1/2 knock-in reveals precision level of PCR tagging. (A,B) N-terminal tagging allows detection of imprecise integration e.g. when indels knock the vamp1/2 coding sequences out of frame, disrupting vesicular localization signals and leading to cytoplasmic FP expression. Examples of F0 larva with likely precise (vesicular) and imprecise (cytoplasmic) mEGFP knock-in into vamp1/2 shown in B. (C) The proportion of vamp1/2 PCR-tagged larvae with likely precise (vesicular) expression pattern is higher when using end-modified HDR templates versus unmodified templates. Number of 3-4 dpf mEGFP+ F0 larva indicated in bars (includes experiments in Fig. 2 and Fig. S4). Error bars indicate 95% CI calculated using the Wilson/Brown method. **P<0.01 or ***P<0.001 (Fisher's test for vesicular versus cytoplasmic/mixed expression). Scale bars: 5 µm.