2′-O-甲氧乙基反义寡核苷酸应用于杜兴肌肉萎缩症的体外研究  

Evaluation of a new antisense oligonucleotide chemistry-2′-O-Methoxyethyl in Duchenne muscular dystrophy in vitro

作　　者：杨璐[1] 王青松[1] 高先军[1] 赵靖雯[1] 韩刚[1] 尹海芳[1] 

机构地区：[1]天津医科大学基础医学研究中心,天津300070

出　　处：《天津医科大学学报》2012年第2期144-147,共4页Journal of Tianjin Medical University

基　　金：国家自然科学基金资助项目(81101340);教育部新世纪优秀人才项目资助(NCET-10-0957)

摘　　要：目的:通过对反义寡核苷酸2′-O-甲氧乙基(2′-O-MOE)化学结构的体外实验,为杜兴肌肉萎缩症基因治疗方法提供一种候选的反义寡核苷酸药物。方法:在杜兴肌肉萎缩症小鼠H2K mdx细胞上转染不同长度和骨架修饰的2′-O-MOE反义寡核苷酸,利用RT-PCR技术检测其诱导外显子跳读的情况。结果:3种2′-O-MOE修饰的反义寡核苷酸体外诱导外显子跳读的效率显著高于目前临床检测的药物2′OmePS,表现出一定的浓度效应和时间效应。并且MOE25(PS)诱导外显子跳读的效率最好。结论:2′-O-MOE修饰的反义寡核苷酸能够有效地诱导H2K mdx细胞外显子跳读。Objective： To study a new antisense oligonucleotide （AO） chemistry- 2＇-O-Methoxyethyl （2＇-O-MOE） in a Duchenne muscular dystrophy （DMD）cellular model- H2K mdx cell and wish to provide a candidate antisense oligonucleotide drug for treating DMD. Methods： RT-PCR was utilized for detecting the exon skipping efficiency after in vitro transfection with different lengths and dif- ferent backbone modifications of 2＇-O-MOE AOs in HEK mdx cells. Results： Three tested 2＇-O-MOE modified AOs showed significant- ly higher level of exon skipping than that of 2＇ OmePS, which was used in current clinical trials. These 2＇-O-MOE modified AOs showed dose-dependent effect and timecourse effect. Among three 2＇-O-MOE modified AOs, MOE25（PS） was the best. Conclusion： It is shown that 2＇-O-MOE modified AOs are effective in inducing exon skipping in vitro.

关 键 词：杜兴肌肉萎缩症 反义寡核苷酸 外显子跳读 

分 类 号：Q5[生物学—生物化学] Q7