The study introduces a novel CRISPR/Cas9-based acetyltransferase fusion protein, dCas9p300 Core, which can activate genes from promoters and enhancers. This fusion protein, consisting of the nuclease-null dCas9 protein fused to the catalytic core of the human acetyltransferase p300, specifically acetylates histone H3 lysine 27 at target sites, leading to robust transcriptional activation. The dCas9p300 Core fusion protein is highly specific and effective in activating genes from enhancer regions and with individual guide RNAs (gRNAs). It also demonstrates the ability to activate genes from distal enhancers and promoters with a single gRNA, simplifying the transactivation process. The p300 Core domain is shown to be portable to other programmable DNA-binding proteins, further enhancing its versatility. The results highlight the potential of this technology in manipulating gene regulation and understanding the relationship between the epigenome and transcriptional control.The study introduces a novel CRISPR/Cas9-based acetyltransferase fusion protein, dCas9p300 Core, which can activate genes from promoters and enhancers. This fusion protein, consisting of the nuclease-null dCas9 protein fused to the catalytic core of the human acetyltransferase p300, specifically acetylates histone H3 lysine 27 at target sites, leading to robust transcriptional activation. The dCas9p300 Core fusion protein is highly specific and effective in activating genes from enhancer regions and with individual guide RNAs (gRNAs). It also demonstrates the ability to activate genes from distal enhancers and promoters with a single gRNA, simplifying the transactivation process. The p300 Core domain is shown to be portable to other programmable DNA-binding proteins, further enhancing its versatility. The results highlight the potential of this technology in manipulating gene regulation and understanding the relationship between the epigenome and transcriptional control.