Braasch Dwaine A, Liu Yinghui, Corey David R
Department of Pharmacology, University of Texas Southwestern Medical Center at Dallas, 5323 Harry Hines Boulevard, Dallas, TX 75390-9041, USA.
Nucleic Acids Res. 2002 Dec 1;30(23):5160-7. doi: 10.1093/nar/gkf651.
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 10 degrees C 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.
使用反义寡核苷酸是实现基因表达调控的一种通用策略。不幸的是，反义诱导表型的解释有时很困难，而改善反义作用效力和特异性的化学修饰将很有用。将锁核酸（LNA）碱基引入寡核苷酸可使结合亲和力得到显著提高，每次取代可达10摄氏度，这使得LNA成为优化反义效力的一个令人兴奋的选择。在这里，我们研究了含LNA碱基的寡核苷酸在细胞内反义基因抑制的规律。含LNA的寡聚物通过阳离子脂质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作用，并鉴定了针对这些位置中每一个的有效反义剂。我们的数据表明，LNA碱基可用于开发反义寡核苷酸，并且它们的使用是一种在细胞内有效抑制基因表达的通用方法。
