Fig. 7

Effects of ZIM3-KRAB domain on CRISPRi-induced tyr knockdown in embryos of X. tropicalis. (A) Schematic illustration of the experimental setup used to analyze the capacity of KRAB domain from Zim3 gene (ZIM3K) to target exogenous reporter gene in X. tropicalis embryos. The gTry13 were injected into the fertilized eggs together with dCas9-KM or dCas9-ZIM3K mRNAs, followed by embryo evaluation at 48 hpi as follows. (B) The qPCR validation of Tyr expression in the injected embryos. Data are presented as mean ± SEM (n = 5 per group). ****p < 0.0001 versus control group (one-way ANOVA test). Ns, no significant differences. (C) Representative images of tyrosinase expression in embryos co-injected with gTry13 and dCas9-mediated repressors at 48 hpi. The top panel shows low magnification (Scale bar = 1 mm), and the lower panel shows high magnification (Scale bar = 500 μm). (D) Embryos with different phenotypes were counted and compared with the total developed ones after injection. Total embryos evaluated for each group (n) is shown above each column