ORIGINAL ARTICLE   

Minerva Biotecnologica 2020 September;32(3):106-13

DOI: 10.23736/S1120-4826.20.02618-X

Copyright © 2020 EDIZIONI MINERVA MEDICA

language: English

Preparation of carbon dot as a potential CRISPR/Cas9 plasmid delivery system for lung cancer cells

Reza MOHAMMADINEJAD ¹, Ali DEHSHAHRI ², Hosseinali SASSAN ³ ✉, Behzad BEHNAM ¹, Milad ASHRAFIZADEH ⁴, Azadeh SAMAREH GHOLAMI ³, Abbas PARDAKHTY ¹, Ali MANDEGARY ¹

¹ Pharmaceutics Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran; ² Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran; ³ Department of Biology, Faculty of Sciences, Shahid Bahonar University, Kerman, Iran; ⁴ Department of Basic Science, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran

BACKGROUND: Nowadays, great attention has been directed to novel gene editing platforms including CRISPR/Cas9 systems. It appears that carbon dots (CDs) might be considered as potential candidates for gene delivery.
METHODS: In the present study, we sought to find an efficient non-viral nanocarrier system to transfer CRISPR/Cas9 plasmid into A549 cell line. The characterization of CDs was performed using transmission electron microscopy, fluorescence emission spectra and zeta (ζ) potential measurement. The delivery potential and biocompatibility of CDs were evaluated using cell uptake (bioimaging) and cell viability assay, respectively.
RESULTS: The CDs had positive charge (+32mV) with spherical shape. The cationic CDs could condense the CRISPR plasmid (pCRISPR) efficiently. Significant viability of the cells following the treatment with CDs demonstrated low toxicity while the CDs could deliver the system into the cells effectively.
CONCLUSIONS: According to our results, cationic CDs have shown potential to act as gene carriers for CRISPR/Cas9 system.

KEY WORDS: Poly(Nepsilon-acryloyllysine); Polyethylenimine; CRISPR-Associated Protein 9; Gene editing