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

Yıl 2022, Cilt: 1 Sayı: 1, 33 - 39, 31.12.2022

Gozde Ultav , Kubra Mac Sena Kizilboga Vedat Gundogdu Hayrettin Tonbul , Emine Şalva

Öz

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.

Anahtar Kelimeler

CRISPR/Cas9, plasmid, silica nanoparticle, delivery system, zeta potential, particle size.

Kaynakça

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