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MiniCAFE, a CRISPR/Cas9-based compact and potent transcriptio
Custom-Made AAVs were used for two-viral-particle injection and all-in-one virus injection experiments. (From BrainVTA)
The viruses used in this article from BrainVTA are in the table below
Custom-Made AAVs  rAAV-gFgf21-CMV-miniCAFE
Xin Zhang, Sihan Lv, Zhenhuan Luo, Yongfei Hu, Xin Peng, Jie Lv, Shanshan Zhao, Jianqi Feng, Guanjie Huang, Qin-Li Wan, Jun Liu, Hongxin Huang, Bing Luan, Dong Wang, Xiaoyang Zhao, Ying Lin, Qinghua Zhou, Zhen-Ning Zhang, Zhili Rong
Pub Date: 2021-03-04, DOI: 10.1093/nar/gkab174, Email: [email protected]
CRISPR-mediated gene activation (CRISPRa) is a promising therapeutic gene editing strategy without inducing DNA double-strand breaks (DSBs). However, in vivo implementation of these CRISPRa systems remains a challenge. Here, we report a compact and robustminiCas9 activator (termed miniCAFE) for in vivo activation of endogenous target genes. The system relies on recruitment of an engineered minimal nuclease-null Cas9 from Campylobacter jejuni and potent transcriptional activators to a target locus by a single guide RNA. It enables robust gene activation in human cells even with a single DNA copy and is able to promote lifespan of Caenorhabditis elegans through activation of longevity-regulating genes. As proof-of-concept, delivered within an all-in-one adeno-associated virus (AAV), miniCAFE can activate Fgf21 expression in the liver and regulate energy metabolism in adult mice. Thus, miniCAFE holds great therapeutic potential against human diseases.

Figure 1. Metabolic regulation by activation of Fgf21 in mouse liver with miniCAFE delivered with an all-in-one AAV vector.
In the current study, based on CjCas9, the authors developed a small and potent transcription activator, miniCAFE, which was able to activate various genes in C. elegans, mice and human cells and induced corresponding phenotypes. Thus, miniCAFE can be a universal tool to activate transcription in a wide spectrum of organisms and holds promise for human disease treatment in the future.
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