CRISPR-Cas9.pptxVIP

Modelling Oncogenic Alterations in Vivo: Adult Animals;1.CRISPR在动物体内的进行基因编辑的优势：;1.1 传统基因编辑技术举例：;2. 用CRISPR-Cas9 编辑特定基因：案例之一;2. 用CRISPR-Cas9 编辑特定基因：案例之二;2. 用CRISPR-Cas9 编辑特定基因：案例之二;;2. 用CRISPR-Cas9 编辑特定基因：案例之三;2. 用CRISPR-Cas9 编辑特定基因：案例之三 ;2. 用CRISPR-Cas9 编辑特定基因：案例之五;Current Challenges and Potential Solutions;concerns delivery of the necessary components to the target cell. Many efforts have been made to increase the efficiency of delivery and to broaden its applicability. Traditional method（first used）：plasmid delivery method Two additional approaches : mRNA and Cas9 ribonucleoproteins (RNP核蛋白) (在体外将sgRNA 与重组Cas9蛋白结合), 可在难转染的细胞中取得很好的效果，但因为RNP转入活体的效率过低，所以很少用它将目标物转入生物体内。;Use of viral vectors seems more practical .These methodologies reach a broad range of cells, especially adult stem cells, which are very difficult to modify using other methods 慢病毒载体： Advantage：Lentiviruses are able to reach almost any cell type drawback： they have mutagenic potential that is intensified when they are combined with designer nucleases（核酸酶） 腺病毒： advantage：具有更强的特异性 drawback：很难将进行广泛使用，因为比慢病毒难生成 腺相关病毒（Adeno-associated viruses ）是最理想的一种转运方式。 Advantage：腺相关病毒是RG1（最安全级别）的基因治疗载体，完全不具有潜在致病性。 drawback：AAV载体容量小，目前最多只能容纳4.7 kb外源DNA片段;Another challenge： ;Types of genome-engineering CRISPR-Cas9 system applications to study cancer.;Summary：;Thanks for listening！