RNA干涉资料.ppt

* * * * * * * * * RNase H是一种核糖核酸内切酶，特异性水解DNA：RNA杂交链上RNA链的磷酸二酯键，产生5核苷酸，该酶对单链的核酸、双链DNA或双链RNA没有作用。 * * * 在这种情况下，RNA干扰并非抑制某一特定的基因，而是触发了眼部正常的免疫响应，才使得血管生长减少。 * * * RNA polymerase II (also called RNAP II and Pol II) is an enzyme found in eukaryotic cells. It catalyzes the transcription of DNA to synthesize precursors of mRNA and most snRNA and microRNA In the process of transcription (by any polymerase) there are three main stages: Initiation; the construction of the RNA polymerase complex on the genes promoter with the help of transcription factors. Elongation; the actual transcription of the majority of the gene into a corresponding RNA sequence, highly moderated by several methods. Termination; the ceasation of RNA transcription and the disassembly of the RNA polymerase complex * * * * Antagomir是经过甲基化、硫代、胆固醇修饰的特殊的miRNA inhibitor，在动物体内具有更高的稳定性和抑制效果 * * * * * * * * * * * * * * * * * * * * * * * * * * Jonathan D Hommel, Robert M Sears, Dan Georgescu, Diana L Simmons Ralph J Dileone Nature Medicine, 2003, 9(12):1539-1544 First disease model created by RNA interference Green – EGFP (infected cells) Red – TH （酪氨酸羟化酶 ） TH knockdown results behavioral deficits Jonathan D Hommel, Robert M Sears, Dan Georgescu, Diana L Simmons Ralph J Dileone Nature Medicine, 2003, 9(12):1539-1544 Pfeifer A., et al. Journal of Clinical Investigation, doi:10.1172/JCI29236 (2006) 当朊病毒蛋白进入动物大脑并抵抗了大脑中的降解机制后，会引起大脑皮层神经元细胞快速退化，引发致命性朊病毒病。朊病毒疾病包括人类CJD、疯牛病等，目前还没有有效治疗措施。一种潜在的治疗方法是，敲除所有出现的朊病毒，包括所有正常的和异常的形式。先前有研究证实敲除小鼠朊病毒基因后，小鼠仍很健康，被灌注scrapie也不会患上朊病毒病。利用RNAi沉默小鼠朊病毒基因中的各种部分，获得多种遗传工程小鼠。结果朊病毒蛋白基因被关闭的细胞的数量达到2/3或者以上数量后，小鼠的寿命显著增长。 与对照组（scrapie感染，存活年限为174天）相比，有65％的细胞朊病毒基因被沉默的小鼠寿命延长了25％。 朊病毒病 siRNA and antisense oligonucleotides Similarities Short(～20 bases) nucleic acids Common methods for delivery to cultured cells Induce post-transcriptional gene silencing by targeting mRNA siRNA and many antisense oligonucleotides cause mRNA cleavage Properties can be altered by introducing modified bases Simil