Calcium-based nanomaterials and their interrelation with chitosan: optimization for pCRISPR delivery
Navid Rabiee, Mojtaba Bagherzadeh, Amir Mohammad Ghadiri, Mahsa Kiani, Sepideh Ahmadi, Vahid Jajarmi, Yousef Fatahi, Abdullah Aldhaher, Mohammadreza Tahriri, Thomas J. Webster, Ebrahim Mostafavi
Abstract
There have been numerous advancements in the early diagnosis, detection, and treatment of genetic diseases. In this regard, CRISPR technology is promising to treat some types of genetic issues. In this study, the relationship between calcium (due to its considerable physicochemical properties) and chitosan (as a natural linear polysaccharide) was investigated and optimized for pCRISPR delivery. To achieve this, different forms of calcium, such as calcium nanoparticles (CaNPs), calcium phosphate (CaP), a binary blend of calcium and chitosan including CaNPs/Chitosan and CaP/Chitosan, as well as their tertiary blend including CaNPs–CaP/Chitosan, were prepared via both routine and green procedures using Salvia hispanica to reduce toxicity and increase nanoparticle stability (with a yield of 85%). Such materials were also applied to the human embryonic kidney (HEK-293) cell line for pCRISPR delivery. The results were optimized using different characterization techniques demonstrating acceptable binding with DNA (for both CaNPs/Chitosan and CaNPs–CaP/Chitosan) significantly enhancing green fluorescent protein (EGFP) (about 25% for CaP/Chitosan and more than 14% for CaNPs–CaP/Chitosan).