Figure 3. Histone acetyltransferase activity partially rescue transcription of NPS strong targets.

(A) Schematic for testing whether recruiting histone acetyltransferase activity can bypass NPS in activating transcription. (B) qPCR analysis showing the relative expression of asb11 and her5, normalized by actb1. When targeted, asb11 and her5 transcription was rescued by dCas9-HAT in MZnps embryos (two-tailed unpaired t-test, ns: not significant, ** P <0.01, *** P <0.001). sgRNAs^asb11 injected embryos were used as a control for her5 expression analysis, and vice versa. sgRNAs^asb11: sgRNAs targeting asb11. sgRNAs^her5: sgRNAs targeting her5. (C) RNA in situ hybridization analysis of asb11 and her5 in wild-type (WT) and MZnps embryos. sgRNA targeting asb11 and her5 were injected separately. (D) Genome tracks showing that accessibility, H3K27ac, and transcription were restored at asb11 and her5 in MZnps embryos by the dCas9-HAT system. Transcription was activated for her11, a gene near her5. Dashed boxes highlight the sgRNA targeting regions. Arrows show regions where H3K27ac is rescued. sgRNA targeting asb11 and her5 were co-injected. (E) Model illustrating the sequence of events of NPS-mediated genome activation. See also Figure S3.