RNA干扰由21和22个核苷酸的RNA介导。
RNA interference is mediated by 21- and 22-nucleotide RNAs.
作者信息
Elbashir S M, Lendeckel W, Tuschl T
机构信息
Department of Cellular Biochemistry, Max-Planck-Institute for Biophysical Chemistry, Am Fassberg 11, D-37077 Göttingen, Germany.
出版信息
Genes Dev. 2001 Jan 15;15(2):188-200. doi: 10.1101/gad.862301.
Abstract
Double-stranded RNA (dsRNA) induces sequence-specific posttranscriptional gene silencing in many organisms by a process known as RNA interference (RNAi). Using a Drosophila in vitro system, we demonstrate that 21- and 22-nt RNA fragments are the sequence-specific mediators of RNAi. The short interfering RNAs (siRNAs) are generated by an RNase III-like processing reaction from long dsRNA. Chemically synthesized siRNA duplexes with overhanging 3' ends mediate efficient target RNA cleavage in the lysate, and the cleavage site is located near the center of the region spanned by the guiding siRNA. Furthermore, we provide evidence that the direction of dsRNA processing determines whether sense or antisense target RNA can be cleaved by the siRNA-protein complex.
摘要
双链RNA（dsRNA）通过一种称为RNA干扰（RNAi）的过程在许多生物体中诱导序列特异性的转录后基因沉默。利用果蝇体外系统，我们证明21和22个核苷酸的RNA片段是RNAi的序列特异性介质。短干扰RNA（siRNA）由长dsRNA通过类似RNase III的加工反应产生。具有突出3'末端的化学合成siRNA双链体在裂解物中介导有效的靶RNA切割，切割位点位于指导siRNA跨越区域的中心附近。此外，我们提供证据表明dsRNA加工的方向决定了正义或反义靶RNA是否能被siRNA-蛋白质复合物切割。
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本文引用的文献
1
Functional anatomy of a dsRNA trigger: differential requirement for the two trigger strands in RNA interference.
Mol Cell. 2000 Nov;6(5):1077-87. doi: 10.1016/s1097-2765(00)00106-4.
2
New insights on homology-dependent silencing of I factor activity by transgenes containing ORF1 in Drosophila melanogaster.
Genetics. 2000 Nov;156(3):1147-55. doi: 10.1093/genetics/156.3.1147.
3
A viral movement protein prevents spread of the gene silencing signal in Nicotiana benthamiana.
Cell. 2000 Sep 29;103(1):157-67. doi: 10.1016/s0092-8674(00)00095-7.
4
Evidence that processed small dsRNAs may mediate sequence-specific mRNA degradation during RNAi in Drosophila embryos.
Curr Biol. 2000 Oct 5;10(19):1191-200. doi: 10.1016/s0960-9822(00)00732-6.
5
A calmodulin-related protein that suppresses posttranscriptional gene silencing in plants.
Science. 2000 Oct 6;290(5489):142-4. doi: 10.1126/science.290.5489.142.
6
The silence of the genes.
Curr Opin Genet Dev. 2000 Oct;10(5):562-7. doi: 10.1016/s0959-437x(00)00128-3.
7
Selective reduction of dormant maternal mRNAs in mouse oocytes by RNA interference.
Development. 2000 Oct;127(19):4147-56. doi: 10.1242/dev.127.19.4147.
8
Human RNase III is a 160-kDa protein involved in preribosomal RNA processing.
J Biol Chem. 2000 Nov 24;275(47):36957-65. doi: 10.1074/jbc.M005494200.
9
dsRNA-mediated gene silencing in cultured Drosophila cells: a tissue culture model for the analysis of RNA interference.
Gene. 2000 Jul 11;252(1-2):95-105. doi: 10.1016/s0378-1119(00)00224-9.
10
Transgene-mediated cosuppression in the C. elegans germ line.
Genes Dev. 2000 Jul 1;14(13):1578-83.
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