Cholaminchloride hydrochloride-cationized gelatin/calcium-phosphate nanoparticles as gene carriers for transgenic chicken production

Abstract Currently, transgenic chickens are a popular method for pharmaceutical protein production. In this study, we employed cationized gelatin/calcium phosphate nanoparticles (ca-GCaPs) with surface modification by cholaminchloride hydrochloride as gene carriers to produce transgenic chickens. To evaluate the transfection efficiency, a plasmid (pEGFP-C1) was encapsulated in the ca-GCaPs for HeLa cells transfection. The ca-GCaPs were applied as gene carriers by direct injection into the area opaca of the chicken embryo blastodisc to produce the transgenic chicken. The particle size of the ca-GCaPs was 350 nm and the zeta potential was +15 mV. Plasmids encapsulated in the ca-GCaPs prevented from DNase I degradation. Based on biocompatibility analysis, the ca-GCaPs produce higher cell viability and less cytotoxicity compared to the commercially available product Lipofectamine™ 2000. The in vitro study indicated that the transfection rate using ca-GCaPs was higher than using cationized gelatin nanoparticles (ca-GPs) because calcium phosphate can provide osmotic pressure to break down lysosomes. In the in vivo study, the egg hatch rate was 100% after ca-GCaPs transfection, and green fluorescence protein expression could be observed in chicken tissue on the fourth day after transfection.