Gene silencing by synthetic U1 Adaptors
Gene silencing by synthetic U1 Adaptors
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DOI：
10.1038/nbt.1525
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发表时间：
2009-03-01
影响因子：
46.9
通讯作者：
Gunderson, Samuel I.
中科院分区：
文献类型：
--
作者：
Goraczniak, Rafal;Behlke, Mark A.;Gunderson, Samuel I.
我们描述了一种基因沉默的方法，采用了不同的反义和RNA干扰的作用机制。U1衔接子是双功能寡核苷酸，具有与靶基因末端外显子中的位点互补的“靶结构域”和与U1小核核糖核蛋白（U1 snRNP）剪接因子的U1小核RNA组分结合的“U1结构域”。U1 snRNP与靶前mRNA的拴系抑制了poly（A）-尾的添加，导致细胞核中RNA种类的降解。U1衔接子可以以序列特异性方式抑制内源基因和报告基因。使用全基因组微阵列分析比较U1衔接子与小干扰RNA（siRNA），表明U1衔接子具有有限的脱靶效应，并且对剪接没有可检测的不良影响。此外，用多个U1衔接子或用U1衔接子和siRNA靶向相同基因强烈增强基因沉默。
We describe a gene silencing method that employs a mechanism of action distinct from those of antisense and RNA interference. U1 Adaptors are bifunctional oligonucleotides with a 'target domain' complementary to a site in the target gene's terminal exon and a 'U1 domain' that binds to the U1 small nuclear RNA component of the U1 small nuclear ribonucleoprotein (U1 snRNP) splicing factor. Tethering of U1 snRNP to the target pre-mRNA inhibits poly(A)-tail addition, causing degradation of that RNA species in the nucleus. U1 Adaptors can inhibit both endogenous and reporter genes in a sequence-specific manner. Comparison of U1 Adaptors with small interfering RNA (siRNA) using a genome-wide microarray analysis indicates that U1 Adaptors have limited off-target effects and no detectable adverse effects on splicing. Further, targeting the same gene either with multiple U1 Adaptors or with a U1 Adaptor and siRNA strongly enhances gene silencing.