Home › Cell Biology › Therapeutic Gene Delivery and Transfection in Human Pancreatic Cancer Cells using Epidermal Growth Factor Receptor-targeted Gelatin Nanoparticles
Cell Biology JoVE (Open Access) Citable · DOI

Therapeutic Gene Delivery and Transfection in Human Pancreatic Cancer Cells using Epidermal Growth Factor Receptor-targeted Gelatin Nanoparticles

DOI: 10.3791/3612-v
What you'll learn
  • ✓ Prepare EGFR-targeted gelatin nanoparticles encapsulating plasmid DNA
  • ✓ Assess transfection efficiency using fluorescence microscopy
  • ✓ Quantify therapeutic gene delivery efficacy in pancreatic cancer cells
Protocol

Type B gelatin-based engineered nanovectors system (GENS) was developed for systemic gene delivery and transfection in the treatment of pancreatic cancer. By modification with epidermal growth factor receptor (EGFR) specific peptide on the surface of nanparticles, they could target on EGFR receptor and release plasmid under reducing environment, such as high intracellular glutathione concentrations.

Difficulty
advanced
Total time
~3-5 days (nanoparticle synthesis, transfection, and analysis)
Model organism
Human pancreatic cancer cells (in vitro)
Biosafety
BSL-1

Steps

1
Prepare plasmid DNA encapsulated EGFR-targeted nanoparticles

Synthesize Type B gelatin-based engineered nanovectors modified with EGFR-specific peptide on the surface and encapsulate plasmid DNA. The nanoparticles are designed to release genetic material in reducing intracellular environments.

▶ 01:29
2
Determine transfection efficiency by fluorescence microscopy

Visualize and quantify successful plasmid transfection in pancreatic cancer cells using fluorescence microscopy imaging to assess nanoparticle-mediated gene delivery.

▶ 05:28
3
Quantify therapeutic efficiency of plasmid-encapsulated nanoparticles

Perform quantitative analysis to measure the therapeutic efficacy of gene delivery and transfection in pancreatic cancer cells treated with the EGFR-targeted nanoparticles.

▶ 07:15
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