核酸的生物合成与降解.pptVIP

RNA interference (RNAi) represents a mechanism invented by nature to protect the genome. In the past few years the field has emerged at a surprisingly high pace. The whole story started when Fire et al. (1998) identified double stranded RNA (dsRNA) as the mediator of gene silencing in C. elegans and referred to the term RNAi. 三. 真核生物转录 4.基因沉默-iRNA What is RNA interference (RNAi) 第三十页，共六十四页，2022年，8月28日 The underlying molecular mechanism of gene silencing provides us with short interfering RNAs (siRNAs) which allows to target any gene with high specificity and efficiency. What is RNA interference (RNAi) 第三十一页，共六十四页，2022年，8月28日 What is the outcome, indeed ? Introduction into cells of a double-stranded RNA corresponding to an expressed gene leads to down-regulation of that gene through degradation of its mRNA 第三十二页，共六十四页，2022年，8月28日 How does the siRNA work ? Introduction of synthetic small double-strand oligonucleotide (21-23 nt length) corresponding to specific gene into cell will specifically knock down corresponding mRNA in the cell 第三十三页，共六十四页，2022年，8月28日 An enzyme complex (Dicer), was discovered in Drosophila (Bernstein et al., 2001) as part of a conserved Dicer family expressed in organisms undergoing RNAi. Dicer contains domains for dsRNA binding, RNA unwinding, and ribonuclease activity, and is associated with additional proteins to drive the cleavage of dsRNA in an ATPdependent manner. The resulting siRNA as part of a multiprotein RNA-inducing silencing complex (RISC) is targeted to the complementary RNA species which is then cleaved. 第三十四页，共六十四页，2022年，8月28日 siRNA: Small interfering RNA 第三十五页，共六十四页，2022年，8月28日 第三十六页，共六十四页，2022年，8月28日 真核基因转录的蛋白因子 为蛋白质编码的基因的准确转录受多种转录因子的调控。RNA聚合酶必须在特定的转录因子的参与下才能起始转录。 至今有20种以上蛋白因子参与转录起始，可分为三类： 通用(基础)转录因子(basal transcriptional activity)： 包括TF II A、TF II B、 TF II D、 TF II E、TF II F、TF II H、TF II J等。这些蛋白因子对于准确转录起始是必需的。 三. 真核生物转录 5.转录因子 第三十七页，共六十四页，2022年，8月28日 2. 启动子特异性转录激活蛋白 (promoter-specific transcriptional