真核基因的转录后调控RNAsilencingsiRNAandmiRNA.PPT

microRNA ta-siRNAs nat-siRNAs ra-siRNAs The Actions Of Small RNAs In Plants Vazquez 2006 Key Points Small RNAs, in the forms of siRNAs and miRNAs, play large roles in the regulation of gene expression in eukaryotes. They are important in normal cell metabolism, development, and defense against invaders. siRNAs are produced from longer segments of dsRNA by Dicer, assembed into RISC, and targeted to mRNAs with perfect complementarity, giving silencing by cleavage and degradation of the RNA or by formation of heterochromatin. The pathway for miRNA has many steps in common with that for siRNA. However, miRNAs are processed from hairpin structures by Drosha and then by Dicer, and they most often have imperfect complementarity with their targets, giving effects on translation rather than Ago-mediated cleavage of the mRNA. 思考题 1，如何判断miRNA在mRNA翻译的哪个阶段（起始、延伸或终止）行使翻译抑制功能？ 2，在Fire等的Nature文章中，他们是怎样想到用dsRNA的？ 3，siRNA的off-target effect 是如何发生的？ 4， 转sense CHS gene为什么会产生红白相间的牵牛花？ Twenty-TveDnucleotide ACO antisense and sense RNA in PTGS lines. (A) Endogenous ACO mRNA abundance in Tve tomato lines containing 35S-ACO transgenes. ACO mRNA was ampliTed by reverse transcriptaseDpolymerase chain reaction and detected by hybridization with labeled ACO cDNA. (B and C) Low molecular weight RNA from the same Tve lines and a 30-nt ACO antisense RNA were fractionated, blotted, and hybridized with either ACO sense RNA (B) or antisense RNA (C) transcribed from full-length ACO cDNA. The low hybridization temperature permitted some nonspeciTc hybridization to tRNA and small ribosomal RNA species, which constitute most of the RNA mass in these fractions. The oligonucleotide hybridized only to the antisense-speciTc probe (B). Twenty-TveDnucleotide, PTGS-speciTc RNA is indicated. * * * (A) Front and side view ribbon representations of Dicer showing the N-terminal platform domain (blue), the PAZ domain (orange), the connector helix (red), the RNase IIIa domain (yellow), the RNase IIIb domain