Goraczniak Rafal, Behlke Mark A, Gunderson Samuel I
Rutgers University, 604 Allison Road, Piscataway, New Jersey 08854, USA.
Nat Biotechnol. 2009 Mar;27(3):257-63. doi: 10.1038/nbt.1525. Epub 2009 Feb 15.
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.
我们描述了一种基因沉默方法，该方法采用的作用机制不同于反义核酸和RNA干扰的作用机制。U1衔接子是双功能寡核苷酸，其“靶结构域”与靶基因末端外显子中的一个位点互补，“U1结构域”则与U1小核核糖核蛋白（U1 snRNP）剪接因子的U1小核RNA成分结合。将U1 snRNP拴系到靶前体mRNA上会抑制聚腺苷酸尾的添加，导致该RNA种类在细胞核中降解。U1衔接子能够以序列特异性方式抑制内源性基因和报告基因。使用全基因组微阵列分析将U1衔接子与小干扰RNA（siRNA）进行比较表明，U1衔接子的脱靶效应有限，并且对剪接没有可检测到的不利影响。此外，用多个U1衔接子或用一个U1衔接子和siRNA靶向同一基因可强烈增强基因沉默。
