MiniCAFE, a CRISPR/Cas9-based compact and potent transcriptional activator, elicits gene expression in vivo

作者：Zhang, Xin; Lv, Sihan; Luo, Zhenhuan; Hu, Yongfei; Peng, Xin; Lv, Jie; Zhao, Shanshan; Feng, Jianqi; Huang, Guanjie; Wan, Qin-Li; Liu, Jun; Huang, Hongxin; Luan, Bing; Wang, Dong; Zhao, Xiaoyang; Lin, Ying; Zhou, Qinghua^*; Zhang, Zhen-Ning^*; Rong, Zhili^*

来源：Nucleic Acids Research, 2021, 49(7): 4171-4185.

DOI：10.1093/nar/gkab174

• GROWTH-FACTOR 21
• C. ELEGANS
• LIFE-SPAN
• GENOMIC LOCI
• HUMAN-CELLS
• CAS9
• CRISPR
• ENDONUCLEASE
• METABOLISM
• PROMOTERS

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 robust miniCas9 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.

• 单位
  同济大学; 南方医科大学