A Versatile Nonviral Delivery System for Multiplex Gene‐Editing in the Liver
Abstract
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.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Oct 14, 2020
- Source ID
- 10.1002/adma.202003537
Entities
People
- Dan Shao
- Dantong Huang
- Hanze Hu
- Hong-Xia Wang
- Jing Gong
- Jonathan Guo
- Kam W. Leong
- Mingqiang Li
- Naazanene Vatan
- Tzu-Chieh Ho
- Yeh-Hsing Lao
Organizations
- Columbia University
- Defense Advanced Research Projects Agency
- Institute of Life Sciences
- National Institutes of Health
- National Science Foundation
- Sun Yat-sen University