Abe T, Takai K, Nakada S, Yokota T, Takaku H
Department of Industrial Chemistry, Chiba Institute of Technology, Tsudanuma, Narashino, Japan.
FEBS Lett. 1998 Mar 20;425(1):91-6. doi: 10.1016/s0014-5793(98)00206-3.
We have designed a new class of oligonucleotides, 'dumbbell RNA/DNA chimeric oligonucleotides', consisting of a sense RNA sequence and its complementary antisense DNA sequence, with two hairpin loop structures. The reaction of the nicked (NDRDON) and circular (CDRDON) dumbbell RNA/ DNA chimeric oligonucleotides with RNase H gave the corresponding antisense phosphodiester oligodeoxynucleotide together with the sense RNA cleavage products. The liberated antisense phosphodiester oligodeoxynucleotide was bound to the target RNA, which gave RNA cleavage products by treatment with RNase H. The circular dumbbell RNA/DNA chimeric oligonucleotide showed more nuclease resistance than the linear antisense phosphodiester oligonucleotide (anti-ODN) and the nicked dumbbell RNA/DNA chimeric oligonucleotide. The CDRDON with four target sites (influenza virus A RNA polymerases (PB1, PB2, PA) and nucleoprotein (NP)) was synthesized and tested for inhibitory effects by a CAT-ELISA assay using the clone 76 cell line. The circular dumbbell DNA/ RNA chimeric oligonucleotide (CDRDON-PB2-as) containing an AUG initiation codon sequence as the target of PB2 showed highly inhibitory effects.
我们设计了一类新型的寡核苷酸，即“哑铃状RNA/DNA嵌合寡核苷酸”，它由一条有义RNA序列及其互补的反义DNA序列组成，并带有两个发夹环结构。带切口的（NDRDON）和环状的（CDRDON）哑铃状RNA/DNA嵌合寡核苷酸与核糖核酸酶H反应，生成相应的反义磷酸二酯寡脱氧核苷酸以及有义RNA的切割产物。释放出的反义磷酸二酯寡脱氧核苷酸与靶RNA结合，经核糖核酸酶H处理后产生RNA切割产物。环状哑铃状RNA/DNA嵌合寡核苷酸比线性反义磷酸二酯寡核苷酸（反义寡核苷酸）和带切口的哑铃状RNA/DNA嵌合寡核苷酸表现出更强的核酸酶抗性。合成了具有四个靶位点（甲型流感病毒RNA聚合酶（PB1、PB2、PA）和核蛋白（NP））的CDRDON，并使用克隆76细胞系通过CAT-ELISA测定法测试其抑制效果。含有作为PB2靶标的AUG起始密码子序列的环状哑铃状DNA/RNA嵌合寡核苷酸（CDRDON-PB2-as）表现出高度抑制作用。
