實驗動物技術應用（一） 基因改造-技術原理 2012/11/02
定義 什麼是基因轉殖？ 利用生物技術改造生物體內的基因 什麼是轉殖動物？ 凡動物具有經分子生物技術改變之基因 如何改造？ 得到基因：轉殖技術 (transgenic) 失去基因：基因剔除技術 (knockout) 複製個體：細胞核轉殖技術 (clone)
Outline Transgenic mice Knockout mice Clone Regulations to production of transgenic animals
小鼠胚胎發育 Blastocyst
Embryonic stem (ES) cells
Transgenic mice Flash DNA Microinjection ES Microinjection Transgenic mice are produced by random integration of a foreign gene into the mouse germ line.
Application Promoter assay Promoter + Reporter gene (Leuciferase, GFP…) Gene product assay Specific Promoter (CMV, SV40) + Gene GFP mice
Mario Capecchi The Nobel Prize in Physiology or Medicine (2007) Knockout mice Mario Capecchi The Nobel Prize in Physiology or Medicine (2007)
Inactivating the Function of Specific Genes in Eukaryotes Molecular Cell Biology 5th Edition
Gene-inactivation Techniques Replacing a normal gene with other sequences; Introducing an allele whose encoded protein inhibits functioning of the expressed normal protein; Promoting destruction of the mRNA expressed from a gene.
Homologous Recombination
Homologous Recombination
Homologous Recombination
Homologous Recombination
Isolation of mouse ES cells with a gene-targeted disruption
Isolation of mouse ES cells with a gene-targeted disruption
ES cells heterozygous for a disrupted gene
Somatic Cell Recombination Can Inactivate Genes in Specific Tissues
Introducing an allele whose encoded protein inhibits functioning of the expressed normal protein
Promoting destruction of the mRNA expressed from a gene- RNA interference (RNAi)
Mario Capecchi The Nobel Prize in Physiology or Medicine (2007) Knockout mice Mario Capecchi The Nobel Prize in Physiology or Medicine (2007)