實驗動物技術應用（一） 基因改造-技術原理

Presentation on theme: "實驗動物技術應用（一） 基因改造-技術原理"— Presentation transcript:

1 實驗動物技術應用（一） 基因改造-技術原理
2012/11/02

2 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)

3 Outline Transgenic mice Knockout mice Clone
Regulations to production of transgenic animals

4 小鼠胚胎發育 Blastocyst

5 Embryonic stem (ES) cells

6 Transgenic mice Flash DNA Microinjection ES Microinjection Transgenic mice are produced by random integration of a foreign gene into the mouse germ line.

7 Application Promoter assay Promoter + Reporter gene (Leuciferase, GFP…) Gene product assay Specific Promoter (CMV, SV40) + Gene GFP mice

8 Mario Capecchi The Nobel Prize in Physiology or Medicine (2007)
Knockout mice Mario Capecchi The Nobel Prize in Physiology or Medicine (2007)

9 Inactivating the Function of Specific Genes in Eukaryotes
Molecular Cell Biology 5th Edition

10 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.

11 Homologous Recombination

12 Homologous Recombination

13 Homologous Recombination

14 Homologous Recombination

15 Isolation of mouse ES cells with a gene-targeted disruption

16 Isolation of mouse ES cells with a gene-targeted disruption

17 ES cells heterozygous for a disrupted gene

18 Somatic Cell Recombination Can Inactivate Genes in Specific Tissues

19 Introducing an allele whose encoded protein inhibits functioning of the expressed normal protein

20 Promoting destruction of the mRNA expressed from a gene- RNA interference (RNAi)

21 Mario Capecchi The Nobel Prize in Physiology or Medicine (2007)
Knockout mice Mario Capecchi The Nobel Prize in Physiology or Medicine (2007)