Department of Biomedical Engineering, Columbia University, New York, NY, 10027, USA.
Laboratory of Biomaterials and Translational Medicine, The Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, 510630, China.
Adv Mater. 2020 Nov;32(46):e2003537. doi: 10.1002/adma.202003537. Epub 2020 Oct 14.
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 genes pcsk9, apoc3, and angptl3 would 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 both pcsk9 and angptl3-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.
CRISPR 的最新进展为基因水平的治疗策略提供了有吸引力的基因组编辑工具。在这里，报道了一种脂质体包覆的介孔硅纳米颗粒（lipoMSN）作为一种有效的 CRISPR 递药系统，可用于肝脏中的多重基因编辑。MSN 可有效负载 Cas9 质粒和 Cas9 蛋白/向导 RNA 核糖核蛋白复合物（RNP），而脂质体包覆则提供了更好的血清稳定性和增强的细胞摄取。假设脂质代谢相关基因 pcsk9、apoc3 和 angptl3 的功能丧失突变通过降低血液胆固醇和甘油三酯来改善心血管健康，该 lipoMSN 用于递送针对这些基因的组合 RNP。在靶向单个基因时，lipoMSN 实现了 54%的基因编辑效率，优于最先进的 Lipofectamine CRISPRMax。对于多重靶向，尽管靶向特定 RNP 的剂量减少，但 lipoMSN 仍能保持每个基因靶标显著的基因编辑效率。通过在同一纳米颗粒中递呈靶向 RNP 的组合，在体外和体内观察到对脂质代谢的协同作用。这些效果，例如在用携带 pcsk9 和 angptl3 靶向 RNP 的 lipoMSN 治疗 4 周后血清胆固醇降低 50%，是单一基因编辑方法无法达到的。总之，这种 lipoMSN 代表了一种多功能平台，可用于开发高效的组合基因编辑疗法。
