BioImpacts : BI · 2026
Saponin conjugation improves p-CRISPR delivery by polyethylenimine modified carbon dots.
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Kelkawi Ali Hamad Abd, Kamrani Mousavi Seyede Mahtab, Hasanzadeh Akbar, Saeedi Sara, Kheiri Yeganeh Azar Behjat, Kamali Babak, Hooshmand Seyyed Emad, Rastgoo Korande Mohadese, Hamblin Michael R, Gaaib Jawdat N, Karimi Mahdi
Аннотация
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.
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.