Antisense inhibition of gene expression in cells by oligonucleotides incorporating locked nucleic acids: effect of mRNA target sequence and chimera design
Antisense inhibition of gene expression in cells by oligonucleotides incorporating locked nucleic acids: effect of mRNA target sequence and chimera design
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DOI：
10.1093/nar/gkf651
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发表时间：
2002-12-01
影响因子：
14.9
通讯作者：
Corey, DR
中科院分区：
文献类型：
--
作者：
Braasch, DA;Liu, YH;Corey, DR
使用反义寡核苷酸是实现基因表达控制的通用策略。不幸的是，反义诱导的表型的解释有时是困难的，并且改善反义作用的效力和特异性的化学修饰将是有用的。将锁核酸（LNA）碱基引入寡核苷酸中可显著提高结合亲和力，每次置换可达10 ℃，使LNA成为优化反义功效的令人兴奋的选择。在这里，我们研究的规则，含有LNA碱基的寡核苷酸在细胞内的反义基因抑制。使用阳离子脂质将含有LNA的寡聚体转染到细胞中并在细胞核中积累。我们测试了与5 '-非翻译区、起始密码子周围区域和mRNA编码区互补的LNA和LNA-DNA嵌合体的反义基因抑制，并鉴定了靶向这些位置中的每一个的有效反义试剂。我们的数据表明，LNA碱基可用于开发反义寡核苷酸，并且它们的使用是有效抑制细胞内基因表达的通用方法。
Use of antisense oligonucleotides is a versatile strategy for achieving control of gene expression. Unfortunately, the interpretation of antisense-induced phenotypes is sometimes difficult, and chemical modifications that improve the potency and specificity of antisense action would be useful. The introduction of locked nucleic acid (LNA) bases into oligonucleotides confers exceptional improvement in binding affinity, up to 10degreesC per substitution, making LNAs an exciting option for the optimization of antisense efficacy. Here we examine the rules governing antisense gene inhibition within cells by oligonucleotides that contain LNA bases. LNA- containing oligomers were transfected into cells using cationic lipid and accumulated in the nucleus. We tested antisense gene inhibition by LNAs and LNA-DNA chimeras complementary to the 5'-untranslated region, the region surrounding the start codon and the coding region of mRNA, and identified effective antisense agents targeted to each of these locations. Our data suggest that LNA bases can be used to develop antisense oligonucleotides and that their use is a versatile approach for efficiently inhibiting gene expression inside cells.