title: Gelatin nanoparticles as gene carriers for transgenic chicken applications.
abstract: To develop a safe and effective nonviral gene delivery system for transgenic chicken manipulation, we developed gelatin nanocarriers using a reporter plasmid (pEGFP-C1; enhanced green fluorescence protein, EGFP) that expressed EGFP. pEGFP-C1-containing gelatin nanoparticles (GP/pEGFP) were prepared using a water-ethanol solvent displacement method and characterized by size, surface charge, DNA loading, and DNA protection ability. For gene delivery, pEGFP-C1 was stably and efficiently encapsulated in GPs that were approximately 300 nm in diameter with a slight negative surface charge, which was prepared from gelatin solution at pH 8.0. Approximately, 85% of the plasmid DNA was encapsulated in the GPs. Electrophoresis results showed that the GPs provided protection against DNase I digestion. We used the GP/pEGFP as a vector to transfect cells and chicken embryos. The vector was nontoxic to cells, and GFP expression was effectively expressed 24 h after HeLa cell transfection. Direct injection was adapted for vector transport to the chicken embryo; injection in the area opaca (Ao) of the egg resulted in the highest hatching rate without affecting embryo development. GFP gene expression in embryo sections was observed 4 days after injection. The results of this study demonstrate that GPs are a suitable nonviral vector for delivering exogenous genes for transgenic chicken manipulation.
authors.affiliation: Division of Medical Engineering Research, National Health Research Institutes, Zhunan Town, Miaoli County 350, Taiwan, ROC.
authors.forename: Ching-Li
authors.initials: CL
authors.lastname: Tseng
authors.forename: Chu-Li
authors.initials: CL
authors.lastname: Peng
authors.forename: Jian-Yuan
authors.initials: JY
authors.lastname: Huang
authors.forename: Jung-Chih
authors.initials: JC
authors.lastname: Chen
authors.forename: Feng-Huei
authors.initials: FH
authors.lastname: Lin
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001672:Biocompatible Materials / Q000737:chemistry; D002642:Chick Embryo; D002645:Chickens / Q000235:genetics*; D005260:Female; D005780:Gelatin / Q000737:chemistry*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008422:Materials Testing; D053769:Nanocapsules / Q000737:chemistry*; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D014162:Transfection / Q000379:methods
chemical_list: D001672:Biocompatible Materials; D053769:Nanocapsules; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D005780:Gelatin