Development of Silica Nanoparticles as a Delivery System for Plasmid-Based Crispr/Cas9

Abstract

Clustered regular interspace short palindromic repeat (CRISPR)/CRISPR-associated system (Cas) is a promising technology for gene editing systems and genome manipulation. Transferring the CRISPR vector to cells is an important aspect of the effective use of this technology. In this study, we aimed to develop a new delivery system using silica nanoparticles (SNPs) with the CRISPR cas9 vector. SNPs were synthesized by the Stöber method. The synthesized nanoparticles were analyzed with the Dynamic Light Scattering (DLS) method and approximately 100 nm SNPs were obtained. EF1a-GFP CRISPR/Cas9 plasmid has been transfected to the Escherichia coli (E.coli) DH5α and isolated from the strain using the plasmid DNA isolation Kit. The isolated pCas-EF1a-GFP CRISPR/Cas9 plasmid was imaged by agarose gel electrophoresis. CRISPR/Cas9 plasmid (pCRISPR) attached to SNP by electrostatic interactions and obtained pCRISPR/SNP complexes were checked by agarose gel electrophoresis. Results show average particle size and zeta potential of obtained pCRISPR/SNP nanoparticles were among 146.6- 272.7 nm and -20.2 - +16,9 mV, respectively and full complexation was achieved at 1/10 pCRISPR/SNP w/w ratio. Consequently, optimized silica nanoparticles can be a good candidate for the delivery of CRISPR/Cas9 plasmid.