Synthesis of Water Soluble Quantum Dots for Monitoring Carrier-DNA Nanoparticles in Plant Cells

Abstract

Fluorescent quantum dots (QDs) have shown great promise for use as biolabels in cell and animal biology and more recently in plant sciences. An important use of QDs is for monitoring the dynamics, intracellular trafficking, and fate of carrier-DNA nanocomplexes in cell transfection and potentially in plant transformation. In this study, a low cost aqueous procedure has been developed to efficiently prepare biocompatible QDs for monitoring nanoparticle-mediated gene transfer in conjunction with molecular breeding of Jatropha curcas. Water-soluble CdSe nanoparticles were synthesized by self-assembly using L-Cysteine as stabilizer and optimal synthesis scheme established by fluorescence spectroscopy. The QDs were used to label chitosan-DNA nanoparticles via electrostatic interaction and the resultant QD-labeled chitosan-DNA complexes were shown to have superior fluorescence properties with red shift of emission and absorption spectra relative to the CdSe QDs alone. This system is being explored as a superior alternative to Agrobacterium-mediated genetic transformation of Jatropha curcas cells. PCR amplification of the full length of the carried reporter gene (GFP) suggests that the DNA was not digested in Jatropha curcas cells transfected with CdSe/CS-DNA complexes. Furthermore, GFP gene expression in the transfected callus cells, as evidenced by fluorescence detection, suggests that the target DNA was integrated into the plant genome.

Gold Functionalized Mesoporous Silica Nanoparticle Mediated Protein and DNA Codelivery to Plant Cells Via the Biolistic Method

“…To date, nanoparticle‐mediated delivery of biogenic molecules to plant cells has been limited to nucleic acids, including double or single stranded DNA3, 24–28 and small interfering RNA 29. Delivery and release of chemical substances such as phenanthrene and plant growth regulators have also been reported 30, 31.…”

Highly efficient uptake of ultrafine mesoporous silica nanoparticles with excellent biocompatibility by Liriodendron hybrid suspension cells

“…However, protoplast-based transformation methods hold disadvantages in that the viability of the protoplasts and their ability to divide are strongly reduced by the chemicals that are applied to disorganize the cell wall. For this reason, recent studies have focused on intact plant cells; it was found to that they are able to achieve endocytosis to directly internalize single-walled carbon nanotubes (CNTs), CdSe/ZnS QDs, or poly (amidoamine) dendrimer from the extracellular environment [14][15][16][17][18]. Other work showed that multi-walled CNTs with attached cellulose are also able to penetrate the cell wall, and transport intracellularly through cellulose-inducing nanoholes [19].…”

Nanobiotechnology in Agricultural Development

“…It has been modifi ed into octapeptide having a condense DNA with high transfection effi ciencies and lower toxicity compared with non-modifi ed chitosan. The integration of DNA through Jatropha curcas cells has been studied through this type of nanoparticles (Wang et al 2011 ) (Table 1 ).…”