后基因组时代的基因调控.ppt

The discovery of miRNAs The first discoered miRNA: lin-4 Breakthrough with BlastN of the second miRNA (stRNA) let-7 Three strategies of miRNA and target recognition (targets are locating in 3’ UTRs). Create induced phenotypes that can be observed over long time spans Create a stably engineered cells can be assayed either in vitro or in vivo, 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. 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. One mechanism of miRNA controlling oncogene expression 微小RNA调控癌基因表达的一种机制。 c-Myc is a helix–loop–helix leucine zipper transcription factor that regulates an estimated 10–15% of genes in the human and Drosophila genomes. c-Myc activates expression of a cluster of six miRNAs on human chromosome 13. (Figure 1) E2F1 is the transcription factor, which is a target of c-Myc that promotes cell cycle progression. Expression of E2F1 is negatively regulated by two miRNAs in this cluster, miR-17-5p and miR-20a. (Figure 1) Used 2’-O-methyl Antisense oligonucleotides to downregulate the level of miR-17-5p and miR-20a, and then analyzed the pr