In C. elegans, high levels of dsRNA allow RNAi in the absence of RDE-4.

In C. elegans, high levels of dsRNA allow RNAi in the absence of RDE-4.
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DOI:
10.1371/journal.pone.0004052
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发表时间:
2008
期刊:
影响因子:
3.7
通讯作者:
Bass, Brenda L.
Bass, Brenda L.
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Habig, Jeffrey W.;Aruscavage, P. Joseph;Bass, Brenda L.

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秀丽隐杆线虫需要一个辅助的双链RNA结合蛋白RDE-4来实施RNA干扰的第一步,即切割dsRNA以产生siRNA。虽然RDE-4通常是RNAi所必需的,但我们报告说,在存在高浓度的触发dsRNA的情况下,RDE-4缺陷的动物能够沉默转基因。在多种标准下,沉默是通过典型的RNAi途径发生的。例如,沉默是RDE-1依赖的,并且随着siRNA的增加而显示出靶向mRNA的减少。我们还发现,高浓度的dsRNA触发导致初级siRNAs的积累增加,这与初级siRNAs向次级siRNAs转换过程中存在限速步骤一致。我们的研究还表明,即使在ADAR存在的情况下,转基因沉默也在胞体中发生在低水平,并且至少有一些siRNAs以依赖于温度的方式积累。我们的结论是,RNAi的反应因条件不同而不同,这可能允许有机体对特定的环境信号做出反应。
C. elegans Dicer requires an accessory double-stranded RNA binding protein, RDE-4, to enact the first step of RNA interference, the cleavage of dsRNA to produce siRNA. While RDE-4 is typically essential for RNAi, we report that in the presence of high concentrations of trigger dsRNA, rde-4 deficient animals are capable of silencing a transgene. By multiple criteria the silencing occurs by the canonical RNAi pathway. For example, silencing is RDE-1 dependent and exhibits a decrease in the targeted mRNA in response to an increase in siRNA. We also find that high concentrations of dsRNA trigger lead to increased accumulation of primary siRNAs, consistent with the existence of a rate-limiting step during the conversion of primary to secondary siRNAs. Our studies also revealed that transgene silencing occurs at low levels in the soma, even in the presence of ADARs, and that at least some siRNAs accumulate in a temperature-dependent manner. We conclude that an RNAi response varies with different conditions, and this may allow an organism to tailor a response to specific environmental signals.
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