武汉大学分子生物学-chapter18-RNA调控.pptVIP

* A mammalian expression vector designed to direct the intracellular synthesis of shRNAs, which will then be processed into siRNAs. * Create induced phenotypes that can be observed over long time spans. Create a stably engineered cells can be assayed either in vitro (体外-细胞水平) or in vivo（体内）, perhaps testing the angiogenic (血管生成) or metastatic (转移) potentials of tumor cells in xenograft models (异种移植模型)。 Create hypomorphic alleles (亚等位基因) rapidly in transgenic mice. [Hypomorphic alleles are those in which function is reduced, but not completely lost] Advantages of expression of siRNA in living cells * 4. Combine shRNAs with existing high-efficiency gene delivery vehicles to create bona fide RNAi-based therapeutics. For example, ultimately, to silence a disease-causing mutant allele specifically. * Research Applications of RNAi: A new strategy of reverse genetics a novel way of gene knock-out It can be used in reverse genetics (反向遗传学) to identify the cellular or biological function of a gene. It can be combined with genomics to perform large-scale genetic screens aimed at gene discovery. * Therapeutic Applications of RNAi:A new strategy to invitation of new drugs and gene therapy siRNAs can be used to counter viral infection by specifically destroying the mRNAs of the pathogenic viruses, such as HIV and HBV. siRNAs can be applied to counter cancers by specifically down-regulate the expression of genes related to oncogenesis. * 本章重点掌握 RNA干扰的发现及其作用机制，微小RNA（miRNA）的发现及其作用机制。 miRNA一些重要的生物学功能（特别是发育），及其在癌症发生中的作用。 siRNA的产生方法及其在科研和治疗上的应用。 * * * * * * Han et al., Cell 125, 887–901, June 2, 2006 * Human Drosha and Dicer share the same RNase III domains and dsRNA binding domain. * 2. MicroRNA (miRNA) targets and regulation. * A comparison between miRNA and siRNA * Three strategies of miRNA and target recognition (targets are locating in 3’ UTRs). * Topic 4: miRNA roles in development, cell differentiation and virus CHAPTER 18 RNAi and miRNA regulation 四、MicroRNA在发育中的调控作用，及其