In vivo targeted delivery of nucleic acids and CRISPR genome editors enabled by GSH-responsive silica nanoparticles

Yuyuan Wang; Pawan K Shahi; Xiuxiu Wang; Ruosen Xie; Yi Zhao; Min Wu; Seth Roge; Bikash R Pattnaik; Shaoqin Gong

doi:10.1016/j.jconrel.2021.06.030

In vivo targeted delivery of nucleic acids and CRISPR genome editors enabled by GSH-responsive silica nanoparticles

J Control Release. 2021 Aug 10:336:296-309. doi: 10.1016/j.jconrel.2021.06.030. Epub 2021 Jun 23.

Authors

Yuyuan Wang¹, Pawan K Shahi², Xiuxiu Wang¹, Ruosen Xie³, Yi Zhao¹, Min Wu⁴, Seth Roge⁴, Bikash R Pattnaik⁵, Shaoqin Gong⁶

Affiliations

¹ Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53715, USA; Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA.
² Department of Pediatrics, University of Wisconsin-Madison, Madison, WI 53706, USA; McPherson Eye Research Institute, University of Wisconsin-Madison, Madison, WI 53705, USA.
³ Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA; Department of Materials Science and Engineering, University of Wisconsin-Madison, Madison, WI 53715, USA.
⁴ Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53715, USA.
⁵ Department of Pediatrics, University of Wisconsin-Madison, Madison, WI 53706, USA; McPherson Eye Research Institute, University of Wisconsin-Madison, Madison, WI 53705, USA; Department of Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, WI 53706, USA.
⁶ Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53715, USA; Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI 53715, USA; McPherson Eye Research Institute, University of Wisconsin-Madison, Madison, WI 53705, USA; Department of Materials Science and Engineering, University of Wisconsin-Madison, Madison, WI 53715, USA; Department of Chemistry, University of Wisconsin-Madison, Madison, WI 53715, USA. Electronic address: shaoqingong@wisc.edu.

Abstract

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.

Keywords: CRISPR-Cas9 genome editing; Gene delivery; Silica nanoparticle.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Animals
CRISPR-Cas Systems
Gene Editing
Glutathione
Mice
Nanoparticles*
RNA, Messenger
Silicon Dioxide*

Substances

RNA, Messenger
Silicon Dioxide
Glutathione

Abstract

Publication types

MeSH terms

Substances

Grants and funding