A Versatile Nonviral Delivery System for Multiplex Gene-Editing in the Liver

作者：Gong, Jing; Wang, Hong-Xia; Lao, Yeh-Hsing; Hu, Hanze; Vatan, Naazanene; Guo, Jonathan; Ho, Tzu-Chieh; Huang, Dantong; Li, Mingqiang; Shao, Dan; Leong, Kam W.^*

来源：Advanced Materials, 2020, 32(46): 2003537.

DOI：10.1002/adma.202003537

• cardiovascular disease
• CRISPR
• Cas9
• gene therapy
• multiplex gene editing
• nanoparticles

Recent advances in CRISPR present attractive genome-editing toolsets for therapeutic strategies at the genetic level. Here, a liposome-coated mesoporous silica nanoparticle (lipoMSN) is reported as an effective CRISPR delivery system for multiplex gene-editing in the liver. The MSN provides efficient loading of Cas9 plasmid as well as Cas9 protein/guide RNA ribonucleoprotein complex (RNP), while liposome-coating offers improved serum stability and enhanced cell uptake. Hypothesizing that loss-of-function mutation in the lipid-metabolism-related genespcsk9,apoc3, andangptl3would improve cardiovascular health by lowering blood cholesterol and triglycerides, the lipoMSN is used to deliver a combination of RNPs targeting these genes. When targeting a single gene, the lipoMSN achieved a 54% gene-editing efficiency, besting the state-of-art Lipofectamine CRISPRMax. For multiplexing, lipoMSN maintained significant gene-editing at each gene target despite reduced dosage of target-specific RNP. By delivering combinations of targeting RNPs in the same nanoparticle, synergistic effects on lipid metabolism are observed in vitro and vivo. These effects, such as a 50% decrease in serum cholesterol after 4 weeks of post-treatment with lipoMSN carrying bothpcsk9andangptl3-targeted RNPs, could not be reached with a single gene-editing approach. Taken together, this lipoMSN represents a versatile platform for the development of efficient, combinatorial gene-editing therapeutics.

• 单位
  中山大学