Nonviral In Vivo Delivery of CRISPR-Cas9 Using Protein-Agnostic, High-Loading Porous Silicon and Polymer Nanoparticles

Abstract

The complexity of CRISPR machinery is a challenge to its application for nonviral in vivo therapeutic gene editing. Here, we demonstrate that proteins, regardless of size or charge, efficiently load into porous silicon nanoparticles (PSiNPs). Optimizing the loading strategy yields formulations that are ultrahigh loading>40% cargo by volumeand highly active. Further tuning of a polymeric coating on the loaded PSiNPs yields nanocomposites that achieve colloidal stability under cryopreservation, endosome escape, and gene editing efficiencies twice that of the commercial standard Lipofectamine CRISPRMAX. In a mouse model of arthritis, PSiNPs edit cells in both the cartilage and synovium of knee joints, and achieve 60% reduction in expression of the therapeutically relevant MMP13 gene. Administered intramuscularly, they are active over a broad dose range, with the highest tested dose yielding nearly 100% muscle fiber editing at the injection site. The nanocomposite PSiNPs are also amenable to systemic delivery. Administered intravenously in a model that mimics muscular dystrophy, they edit sites of inflamed muscle. Collectively, the results demonstrate that the PSiNP nanocomposites are a versatile system that can achieve high loading of diverse cargoes and can be applied for gene editing in both local and systemic delivery applications.

The Current Situation and Development Prospect of Whole-Genome Screening

“…On the other hand, the difficult is that LNPs can be exocytosed. Recently, an increase in the number of modified nanoparticles has been reported to increase the delivery efficiency for the widespread application of gene editing, which provides hope for basic research and the clinical treatment of various diseases [ 98 , 99 , 100 , 101 , 102 , 103 , 104 , 105 , 106 , 107 , 108 , 109 , 110 ].…”

Polyurethane Nanoparticles as Versatile Tools in Nanomedicine: A Review

“…Nanomedicine is currently at an early stage, but it is expected to have a revolutionary impact on health care . In comparison to conventional drugs, nanomedicines have the potential to be more effective, bioavailable, dose-responsive, targetable, personalized, and safe. − Polymer nanoparticles have recently come to be recognized as crucial tools in the field of nanomedicine, particularly for medical imaging, concurrent medication delivery and diagnosis, and therapeutic delivery. − They offer a unique combination of versatility, biocompatibility, and tunability . Nanoparticles have unique properties like high dynamic stability, an extensive range of structural characteristics, diversity, rigid morphology, the ability to encapsulate various hydrophobic or hydrophilic drugs, and an abundance of modification options.…”