Application of chitosan/DNA nanoparticles in transgenic chicken

• Main Authors: Chia-Cheng Li, 李家成
• Other Authors: 林峰輝
• Format: Others
• Language: zh-TW
• Published: 2005
• Online Access: http://ndltd.ncl.edu.tw/handle/51767234303344083434

碩士 === 國立臺灣大學 === 醫學工程學研究所 === 93 === In this study, preparation of well-dispersed chitosan/DNA nanoparticles with size of 100-200 nm was developed. By ionic gelation between chitosan and sodium tripolyphosphate, chitosan nanoparticles with size below 100 nm were prepared under adequate concentration. Furthermore, the mechanism of chitosan nanoparticles formation was expounded by nucleation theory and zetapotential phenomenon. Chitosan/DNA by 10/1 in mass ratio, DNA could be encapsulated effectively in the nanoparticles; meanwhile, size analysis was performed under dynamic light scattering, TEM, and SEM. The stability of the nanoparticles was proven by zetapential analysis. DNA encapsulated in the nanoparticles could be protected from DNAseⅠ degradation. In 293 HEK cells transfection experiment, efficiencies of chitosan/DNA nanoparticles and lipofectamineTM 2000-DNA were 47.53 % and 87.8 % respectively. The cell viability of lipofectamineTM 2000-DNA was much lower than that of chitosan/DNA nanoparticles. The egg hatchability after chitosan/DNA nanoparticles transfection was far better than that of lipofectamineTM 2000-DNA transfection one. Although genomic DNA was not detected in all chicks, if the advantage of good biocompatibility of chitosan/DNA nanoparticles is cooperated with other powerful transgenic techniques or strategies, chitosan/DNA nanoparticles will have great potential in generating transgenic chicken.