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Polyethylenimine-Functionalized Carbon Dots for Delivery of Crispr/Cas9 Complexes Publisher Pubmed



Hashemzadeh I1, 2, 3 ; Hasanzadeh A1, 2, 3 ; Radmanesh F4, 5 ; Khodadadi Chegeni B3 ; Hosseini ES1, 2 ; Kiani J6, 7 ; Shahbazi A8 ; Naseri M6, 7 ; Fatahi Y9, 10, 11 ; Nourizadeh H3 ; Kheiri Yeghaneh Azar B7, 12 ; Aref AR13 ; Liu Y14, 15 ; Hamblin MR16, 17 Show All Authors
Authors
  1. Hashemzadeh I1, 2, 3
  2. Hasanzadeh A1, 2, 3
  3. Radmanesh F4, 5
  4. Khodadadi Chegeni B3
  5. Hosseini ES1, 2
  6. Kiani J6, 7
  7. Shahbazi A8
  8. Naseri M6, 7
  9. Fatahi Y9, 10, 11
  10. Nourizadeh H3
  11. Kheiri Yeghaneh Azar B7, 12
  12. Aref AR13
  13. Liu Y14, 15
  14. Hamblin MR16, 17
  15. Karimi M1, 2, 3, 6, 18, 19
Show Affiliations
Authors Affiliations
  1. 1. Cellular and Molecular Research Center, Iran University of Medical Sciences, Tehran, 1449614535, Iran
  2. 2. Department of Medical Nanotechnology, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, 1449614535, Iran
  3. 3. Advanced Nanobiotechnology and Nanomedicine Research Group (ANNRG), Iran University of Medical Sciences, Tehran, 1449614535, Iran
  4. 4. Uro-oncology Research Center, Tehran University of Medical Sciences, Tehran, 1417613151, Iran
  5. 5. Department of Cell Engineering, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, Tehran, 1665659911, Iran
  6. 6. Oncopathology Research Center, Iran University of Medical Sciences, Tehran, 1449614535, Iran
  7. 7. Department of Molecular Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, 1449614535, Iran
  8. 8. Department of Neuroscience, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, 1449614535, Iran
  9. 9. Nanotechnology Research Centre, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, 1417613151, Iran
  10. 10. Department of Pharmaceutical Nanotechnology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, 1417613151, Iran
  11. 11. Universal Scientific Education and Research Network (USERN), Tehran, 1419733151, Iran
  12. 12. Student Research Committee, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, 1449614535, Iran
  13. 13. Belfer Center for Applied Cancer Science, Department of Medical Oncology, Harvard Medical School, Boston, 02215, Massachusetts, United States
  14. 14. Eng. Res. Ctr. of Clin. Funct. Materials and Diagnosis and Treatment Devices of Zhejiang Province, Wenzhou Institute, University of Chinese Academy of Sciences, Wenzhou, 325000, Zhejiang, China
  15. 15. Oujiang Laboratory, Wenzhou, 325000, Zhejiang, China
  16. 16. Laser Research Centre, Faculty of Health Science, University of Johannesburg, Doornfontein, 2028, South Africa
  17. 17. Radiation Biology Research Center, Iran University of Medical Sciences, Tehran, 1449614535, Iran
  18. 18. Research Center for Science and Technology in Medicine, Tehran University of Medical Sciences, Tehran, 1417613151, Iran
  19. 19. Applied Biotechnology Research Centre, Tehran Medical Science, Islamic Azad University, Tehran, 1916893813, Iran

Source: ACS Applied Bio Materials Published:2021


Abstract

Carbon dots (CDs) have become the focus of many studies due to their outstanding optical properties and good biocompatibility. We investigated their potential application to produce a smart and highly efficient yet nontoxic nanovector for gene delivery. This was achieved by conjugating PEI1.8k-functionalized CDs (synthesized by one-step microwave-assisted pyrolysis) with arginine-disulfide linkers to produce CD-PEI1.8k-Arg nanoparticles. This nanovector could deliver p-CRISPR (9.3 kb) into different types of cell lines with higher efficiency compared to native PEI1.8k or PEI25k. CD-PEI1.8k-Arg also maintained its outstanding transfection efficiency at a high serum concentration and low p-CRISPR dose, compared to PEI25k, which was ineffective under those conditions. Additionally, CD-PEI1.8k-Arg could knock out the GFP gene with great efficiency by delivering the required components of CRISPR/Cas9, including a plasmid encoding Cas9, sgRNA targeting GFP, and Cas9/sgRNA ribonucleoproteins (RNPs) into the HEK 293T-GFP cells. Moreover, the nanoparticles showed potential for the local delivery of p-CRISPR into brain tissue. The remarkable properties of CD-PEI1.8k-Arg could enable the development of a safe, highly efficient gene-delivery nanovector for the treatment of various diseases in the near future. © 2021 American Chemical Society. All rights reserved.
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