Gene Transfection Efficiency Improvement with Lipid Conjugated Cationic Carbon Dots

ACS Appl Mater Interfaces. 2024 May 29;16(21):27087-27101. doi: 10.1021/acsami.4c02614. Epub 2024 May 16.

Abstract

An ideal vehicle with a high transfection efficiency is crucial for gene delivery. In this study, a type of cationic carbon dot (CCD) known as APCDs were first prepared with arginine (Arg) and pentaethylenehexamine (PEHA) as precursors and conjugated with oleic acid (OA) for gene delivery. By tuning the mass ratio of APCDs to OA, APCDs-OA conjugates, namely, APCDs-0.5OA, APCDs-1.0OA, and APCDs-1.5OA were synthesized. All three amphiphilic APCDs-OA conjugates show high affinity to DNA through electrostatic interactions. APCDs-0.5OA exhibit strong binding with small interfering RNA (siRNA). After being internalized by Human Embryonic Kidney (HEK 293) and osteosarcoma (U2OS) cells, they could distribute in both the cytoplasm and the nucleus. With APCDs-OA conjugates as gene delivery vehicles, plasmid DNA (pDNA) that encodes the gene for the green fluorescence protein (GFP) can be successfully delivered in both HEK 293 and U2OS cells. The GFP expression levels mediated by APCDs-0.5OA and APCDs-1.0OA are ten times greater than that of PEI in HEK 293 cells. Furthermore, APCDs-0.5OA show prominent siRNA transfection efficiency, which is proven by the significantly downregulated expression of FANCA and FANCD2 proteins upon delivery of FANCA siRNA and FANCD2 siRNA into U2OS cells. In conclusion, our work demonstrates that conjugation of CCDs with a lipid structure such as OA significantly improves the gene transfection efficiency, providing a new idea about the designation of nonviral carriers in gene delivery systems.

Keywords: cationic carbon dots; gene delivery; lipid; plasmid DNA; small interfering RNA.

MeSH terms

  • Carbon* / chemistry
  • Cations / chemistry
  • Cell Line, Tumor
  • DNA / chemistry
  • Gene Transfer Techniques
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • HEK293 Cells
  • Humans
  • Lipids / chemistry
  • Oleic Acid / chemistry
  • Quantum Dots / chemistry
  • RNA, Small Interfering* / chemistry
  • RNA, Small Interfering* / metabolism
  • Transfection* / methods

Substances

  • Carbon
  • RNA, Small Interfering
  • Lipids
  • Cations
  • DNA
  • Oleic Acid
  • Green Fluorescent Proteins