必威体育精装版基因组编辑技术CRISPRCAS_system课件.pptx

Liang Jiwang2015.02.02Fighting invasion. When viruses (green) attack bacteria, the bacteria respond with DNA-targeting defenses that biologists have learned to exploit for genetic engineering.Lactic acidLactoseInvasion. Streptococcus thermophilus (嗜热链球菌) such as this one may acquire key defensive sequence from infectious bacteriophage, attached at top. 发现Clustered Regularly Interspaced Short Palindromic Repeats免疫过程PAM: proto-spacer adjacent motifs(保守的间隔相邻基序)crRNA: CRISPR RNAtracrRNA: trans-activating CRISPR RNA人工改造PAM(一般为NGG)工作原理HDR: Homology-direct repair 同源直接修复NHEJ : Non-homologous end joining 非同源性末端接合工作原理-切割后的处理HRMA:High-resolution melting analysis(高分辨率熔解曲线分析)A. Bassett, J.-L. Liu / Methods 69 (2014) 128–136应用— 果蝇yellow genewhite geneA. Bassett, J.-L. Liu / Methods 69 (2014) 128–136应用— 果蝇清除感染细胞中的HIVDeletion(190 bp)LTR: long terminal repeats/pubmedroc Natl Acad Sci U S A.?2014 Aug 5;111(31)应用— HIV预防再度感染ICC: Immunocytochemistry免疫细胞化学技术WB: Western blotMOI: multiplicity of infection 感染复数/pubmedroc Natl Acad Sci U S A.?2014 Aug 5;111(31)应用— HIVDrug-target interactionXPO-1/pubmed/?term=Identifying+Drug-Target+Selectivity+of+Small-Molecule+CRM1/XPO1+Inhibitors+by+CRISPR/Cas9+Genome+EditingChem Biol.?2015 Jan 22;22(1):107-16应用— 医药应用— 医药应用— 激活基因Precise cuts. In just 8 months, CRISPR modifications of DNA resulted in dumpier nematodes (left), zebrafish embryos with an excess of ventral tissue (middle), and fruit flies with dark eyes (right), demonstrating its broad utility for editing genes in animals.