A proof-of-concept will be identified for the loading of the plasmid DNA on quantum dots. The quantum dots will be treated with compounds to develop designer functional groups that will help in coating or adsorption of plasmid DNA on the surface of the Qdots. The loading of the DNA molecule on Qdots will be identified through gel electrophoresis analysis. The plasmid DNA-Qdot Nano-complex will then be incubated with the callus tissue suspension derived from different explants for delivery of the target plasmid DNA or reporter gene. The immature embryos extracted/ obtained from the wheat inflorescence will also be incubated with the plasmid DNA-Qdot Nano-complex. The comparative transformation efficiencies for both the callus suspension and immature embryos will be evaluated. The occurrence of the plasmid DNA will be validated through PCR technique. The unloading of the plasmid DNA payload will also be determined through fluorescence microscopy or fluorospectroscopy techniques. The change in the fluorescence emission signal of the Qdots pre and post unloading of the plasmid DNA will be quantified. The Nano-delivery of the plasmid DNA coding for the Cas9 endonuclease and single guide RNA will be performed in callus tissue/ immature embryo. The transformation and genome editing efficiencies will then be determined by performing the T-Endonuclease Assay.
