Ali Hamad Abd Kelkawi
1,2 
, Seyede Mahtab Kamrani Mousavi
3,4,5, Akbar Hasanzadeh
3,4,5, Sara Saeedi
3,4,5, Behjat Kheiri Yeganeh Azar
6,7, Babak Kamali
3,4,5, Seyyed Emad Hooshmand
3,4,5, Mohadese Rastgoo Korande
5, Michael R Hamblin
8, Jawdat N. Gaaib
9,10, Mahdi Karimi
3,4,5,11,10,12* 1 College of Nursing, University of Al-Ameed, Karbala, Iraq
2 Department of Medical Physics, College of Applied Medical Sciences, University of Kerbala, Iraq
3 Cellular and molecular Research Center, Iran University of Medical Sciences, Tehran, Iran
4 Department of Medical Nanotechnology, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran
5 Advanced Nanobiotechnology and Nanomedicine Research Group (ANNRG), Iran University of Medical Sciences, Tehran, Iran
6 Department of Molecular Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran
7 Student Research Committee, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran
8 Laser Research Center, Faculty of Health Science, University of Johannesburg, Doornfontein 2028, South Africa
9 Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, University of Kerbala, Karbala, Iraq
10 University of Al-Ameed, Karbala, Iraq
11 Research Center for Science and Technology in Medicine, Tehran University of Medical Sciences, Tehran, Iran
12 Applied Biotechnology Research Centre, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran
Abstract
Introduction: Non-toxic and efficient delivery platforms are important for gene editing applications. In this report, the preparation of saponin-conjugated low molecular weight polyethyleneimine (PEI1.8kDa) modified carbon dots (CD-PEI-Sap) is described.
Methods: Polyethyleneimine modified carbon dots (CD-PEI1.8kDa), were synthesized by one-step microwave-mediated pyrolysis of citric acid (CA) and PEI1.8kDa to produce non-toxic fluorescent particles. When saponin was added at this stage followed by microwave treatment a completely clear solution of CD-PEI-Sap was produced.
Results: These 80 nm nanoparticle vectors not only transfected green fluorescent protein plasmid (p-GFP) (3 kb) with 60% expression in human embryonic kidney (HEK 293T) cells, and > 60% in hard to transfect human breast adenocarcinoma cells (MCF7), but could also transfect a large clustered regularly interspaced short palindromic repeat plasmid (p-CRISPR) with 5-fold higher efficiency (50%) than the gold standard reagent, branched PEI MWt 25 kDa (PEI25kDa). Moreover, the CD-PEI-Sap system produced good transfection with a low concentration of p-CRISPR and in the presence of 30% fetal bovine serum (FBS).
Conclusion: This system demonstrated efficient transfection and serum resistance, making it a promising tool for future gene editing research.