Plant bio-transformable HMG-CoA reductase gene loaded calcium phosphate nanoparticle: in vitro characterization and stability study

Curr Drug Discov Technol. 2013 Mar;10(1):25-34. doi: 10.2174/1570163811310010005.

Abstract

Encapsulation of plasmid DNA in nanoparticle is expected to enhance the stability of DNA, reproducibility and frequency of the genetic transformation in plants. Here we report the formulation of HMG Co-A reductase gene loaded calcium phosphate nanoparticles (Cap nanoparticles) and their in-vitro, in-vivo characterization. The developed Cap nanoparticles were characterized by DSC, FT-IR, and XRD. Developed Cap nanoparticles were spherical in shape having the particle size and zeta potential in the range of 10.86±0.09nm to 33.42±0.18nm and -25.5±0.07mV to -31.7±0.07mV (for Cap-I to Cap-IV). DNA releasing in acidic media showed, initially slow release followed by fast release with a maximum release of Cap-I (95.77±1.39%) > Cap-II (87.32±2.07%) > Cap-III (76.54±2.01%) > Cap-IV (72.93±1.75%) over 60min. Cap nanoparticles were quite stable at storage condition of 40±0.5°C/75±5%RH, 25±0.5°C/60±RH, 4±0.5°C/ambient humidity and the integrity of pDNA encapsulated was confirmed by gel electrophoresis. Compared to wild type C. intybus, transformation efficiency and enhanced biosynthesis of esculin with the DNA nanoparticles in C. intybus were about 10% and 71%, respectively. Antioxidant activity capacity of the biotransformed plants was significantly higher than the normal plant due to high accumulation of esculin.

MeSH terms

  • Antioxidants / pharmacology
  • Calcium Phosphates / chemistry*
  • Cichorium intybus / genetics*
  • Cichorium intybus / metabolism
  • Drug Carriers / chemistry*
  • Drug Stability
  • Esculin / metabolism
  • Gene Transfer Techniques*
  • Hydroxymethylglutaryl CoA Reductases / genetics*
  • Nanoparticles / chemistry*
  • Plant Extracts / pharmacology
  • Plant Leaves
  • Plants, Genetically Modified

Substances

  • Antioxidants
  • Calcium Phosphates
  • Drug Carriers
  • Plant Extracts
  • Esculin
  • calcium phosphate
  • Hydroxymethylglutaryl CoA Reductases