通过反义寡核苷酸修复哺乳动物细胞中的地中海贫血人类β-珠蛋白mRNA
Repair of thalassemic human beta-globin mRNA in mammalian cells by antisense oligonucleotides.
作者信息
Sierakowska H, Sambade M J, Agrawal S, Kole R
机构信息
Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill 27599, USA.
出版信息
Proc Natl Acad Sci U S A. 1996 Nov 12;93(23):12840-4. doi: 10.1073/pnas.93.23.12840.
Abstract
In one form of beta-thalassemia, a genetic blood disorder, a mutation in intron 2 of the beta-globin gene (IVS2-654) causes aberrant splicing of beta-globin pre-mRNA and, consequently, beta-globin deficiency. Treatment of mammalian cells stably expressing the IVS2-654 human beta-globin gene with antisense oligonucleotides targeted at the aberrant splice sites restored correct splicing in a dose-dependent fashion, generating correct human beta-globin mRNA and polypeptide. Both products persisted for up to 72 hr posttreatment. The oligonucleotides modified splicing by a true antisense mechanism without overt unspecific effects on cell growth and splicing of other pre-mRNAs. This novel approach in which antisense oligonucleotides are used to restore rather than to down-regulate the activity of the target gene is applicable to other splicing mutants and is of potential clinical interest.
摘要
在一种遗传性血液疾病——β地中海贫血的一种形式中，β珠蛋白基因内含子2（IVS2-654）的突变导致β珠蛋白前体mRNA的异常剪接，进而导致β珠蛋白缺乏。用针对异常剪接位点的反义寡核苷酸处理稳定表达IVS2-654人β珠蛋白基因的哺乳动物细胞，可呈剂量依赖性地恢复正确剪接，产生正确的人β珠蛋白mRNA和多肽。两种产物在处理后可持续长达72小时。这些寡核苷酸通过真正的反义机制改变剪接，而对细胞生长和其他前体mRNA的剪接没有明显的非特异性影响。这种使用反义寡核苷酸来恢复而非下调靶基因活性的新方法适用于其他剪接突变体，具有潜在的临床意义。
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