There are very interesting materials chemistry challenges in the development of nanovectors for wheat research because the cell walls and prominent exine in microspores makes endocytosis and subsequent delivery of protein/DNA cargo much more difficult than in mammalian cells. Silica nanocarriers are promising candidates due to their multifunctionality, stability, tunable size and charge and biocompatibility. In this project, our objective is to develop a series of multifunctional luminescent silica nanoparticles that are surface-modified with a variety of cationic ligands/cell penetrating peptides and subsequently use to immobilize gene editing tools/plasmid DNA/ gene cassettes for delivery to wheat microspores. The luminescence built into the nanoparticles provides the unique ability to track the uptake of the carriers into the microspores.
Here, we prepared complexes of plasmid DNA (pDNA, labeled and unlabeled) and silica nanoparticles (SNP) functionalized with amine crosslinked fluorescein isothiocyanate fluorophore (SNP-FITC-APTES) through electrostatic interaction. The resulting SNP/pDNA complexes were isolated/ purified and then the optimized ratios were used for transfection studies into wheat microspores. The preliminary results indicate the transfection of plasmid DNA into wheat microspores through designed SNP/pDNA nanovectors.