The application of chitosan-UA/DNA nanoparticles as gene carrier in transgenic chicken

• Main Authors: Yi-Cheng Tseng, 曾意誠
• Other Authors: 林峰輝
• Format: Others
• Language: zh-TW
• Online Access: http://ndltd.ncl.edu.tw/handle/10019491876015623050

碩士 === 國立臺灣大學 === 醫學工程學研究所 === 94 === There are many therapeutic proteins and peptide have been licensed for production using bacterial, fungal and mammalian cells, and many therapeutic proteins are currently being developed and tested in a variety of host cells. Transgenic animals describes animals with chromosomes that contain stably integrated copies of genes. The use of transgenics as a technology for producing recombinant proteins has made remarkable strides in the past few years. Recently, transgenic chickens should be near-perfect bioreactors for making large amounts of pure recombinant proteins. In this study, we tried to use urocanic acid to modify chitosan as a new vector in gene delivery. We made the nanoparticles with diameter is 150~300 nm, and the DNA loading efficiency is over 90 %. When N/P=2, the zeta potential is +9 mV. And the modified chitosan/DNA nanoparticles can protect the DNA efficiently than unmodified chitosan/DNA. The modified chitosan had very good biocompatibility in 3T3 and HeLa, but the transfection efficiency is cell-dependant. In vivo test, the viability of the treated chicken embryos was down to 14%, and no any green fluroence protein was observed. But the modified chitosan/DNA nanoparticle still has a lot of advantages which applied in transgenic chicken.