Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53715, USA; Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA.
Department of Pediatrics, University of Wisconsin-Madison, Madison, WI 53706, USA; McPherson Eye Research Institute, University of Wisconsin-Madison, Madison, WI 53705, USA.
J Control Release. 2021 Aug 10;336:296-309. doi: 10.1016/j.jconrel.2021.06.030. Epub 2021 Jun 23.
The rapid development of gene therapy and genome editing techniques brings up an urgent need to develop safe and efficient nanoplatforms for nucleic acids and CRISPR genome editors. Herein we report a stimulus-responsive silica nanoparticle (SNP) capable of encapsulating biomacromolecules in their active forms with a high loading content and loading efficiency as well as a well-controlled nanoparticle size (~50 nm). A disulfide crosslinker was integrated into the silica network, endowing SNP with glutathione (GSH)-responsive cargo release capability when internalized by target cells. An imidazole-containing component was incorporated into the SNP to enhance the endosomal escape capability. The SNP can deliver various cargos, including nucleic acids (e.g., DNA and mRNA) and CRISPR genome editors (e.g., Cas9/sgRNA ribonucleoprotein (RNP), and RNP with donor DNA) with excellent efficiency and biocompatibility. The SNP surface can be PEGylated and functionalized with different targeting ligands. In vivo studies showed that subretinally injected SNP conjugated with all-trans-retinoic acid (ATRA) and intravenously injected SNP conjugated with GalNAc can effectively deliver mRNA and RNP to murine retinal pigment epithelium (RPE) cells and liver cells, respectively, leading to efficient genome editing. Overall, the SNP is a promising nanoplatform for various applications including gene therapy and genome editing.
基因治疗和基因组编辑技术的快速发展提出了开发安全有效的核酸和 CRISPR 基因组编辑纳米平台的迫切需求。在此，我们报告了一种刺激响应型二氧化硅纳米颗粒（SNP），它能够以高载药含量和载药效率以及可控的纳米颗粒尺寸（约 50nm）将生物大分子封装在其活性形式中。二硫键交联剂被整合到二氧化硅网络中，当被靶细胞内化时，赋予 SNP 谷胱甘肽（GSH）响应的货物释放能力。将含咪唑的成分掺入 SNP 中，以增强内涵体逃逸能力。SNP 可以高效且具有生物相容性地递送各种货物，包括核酸（如 DNA 和 mRNA）和 CRISPR 基因组编辑物（如 Cas9/sgRNA 核糖核蛋白（RNP）和带有供体 DNA 的 RNP）。SNP 表面可以聚乙二醇化并通过不同的靶向配体进行功能化。体内研究表明，与全反式视黄酸（ATRA）缀合的视网膜下注射 SNP 和与 GalNAc 缀合的静脉内注射 SNP 可以分别有效地将 mRNA 和 RNP 递送至小鼠视网膜色素上皮（RPE）细胞和肝细胞，从而实现有效的基因组编辑。总的来说，SNP 是一种很有前途的纳米平台，可用于包括基因治疗和基因组编辑在内的各种应用。
