FITC Labelled Poly (?-Amino Ester) Nanoparticles For Using Sperm Mediated Gene Transfer
5th International Group Congress – “Vet-Istanbul Group” & 8th International Scientific Meeting “Days of Veterinary Medicine-2018, Ohrid, Macedonia, 23 - 27 September 2018, vol.1, no.1, pp.127, (Full Text)
- Publication Type: Conference Paper / Full Text
- Volume: 1
- City: Ohrid
- Country: Macedonia
- Page Numbers: pp.127
- Istanbul University Affiliated: Yes
Abstract
Abstract
The transgenic animals have a purposeful modification on their genome (DNA). It is aimed to understand the gene function of the human disease and consider the effects of therapeutic intervention and the other aim is to manufacture great amount of complex human protein or organ donor. There are many advanced biotechnological methods for transgenic animal production. One of them is Sperm Mediated Gene Transfer (SMGT) which using the spermatazoa as a natural vector for gene carrier. It helps avoiding of immunological response, whereas there are unexpected interaction between spermatozoa and desired gene construct in the last researches. There are natural and synthetic cationic biopolymers used as gene delivery systems. Poly (β-amino ester)s (PBAEs) which are one of the most popular synthetic biopolymers, have attraction as efficient, biodegradable and low cytotoxity. PBAEs as cationic biopolymers are capable of binding and condensation to negatively charged DNA into stable nanoparticles. In this study, PBAE was synthesized via the Michael addition between the diacrylate and the diamine monomers. PBAE nanoparticles were prepared with solvent diffusion technique and fluorescein isothiocyanate (FITC) labelled PBAE nanoparticles (PBAE-FITC) were prepared for SMGT technique. The content of the FITC of the PBAE-FITC was also determined using by UV/vis spectroscopy. The physical properties of the PBAE-FITC were determined and found to be the optimum particle size of 90,42 nm, the polydispersity index (PDI) of 0,100 and the zeta potential of 32,1 mV. The results of the binding efficacy of PBAE-FITC to spermatozoa’s membrane were also evaluated using by flow cytometry. As a result of the preliminary in vitro studies, it was observed that the 81,64 % of spermatozoa were labelled with PBAE-FITC.
Acknowledgements: This study was funded by Istanbul University with the project number: 28733