Woolf T M, Melton D A, Jennings C G
Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, MA 02138.
Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7305-9. doi: 10.1073/pnas.89.16.7305.
Antisense oligonucleotides are widely used as inhibitors of gene expression in cultured cells and have been proposed as potential therapeutic agents, but it is not known to what extent they are specific for their intended target RNAs. Statistical considerations indicate that if oligonucleotides can form hybrids with mRNA molecules in vivo by means of short or imperfect regions of complementarity, then the specificity of oligonucleotides as antisense reagents will be greatly compromised. We have used Xenopus oocytes as a model system in which to investigate the potential specificity of antisense oligonucleotides in vivo. We injected perfect and partially matched antisense oligonucleotides into oocytes and measured the resulting degradation of the target RNA in each case. On the basis of the extent to which antisense oligonucleotides can cause cleavage of RNAs at imperfectly matched target sites, we conclude that in this system it is probably not possible to obtain specific cleavage of an intended target RNA without also causing at least the partial destruction of many nontargeted RNAs.
反义寡核苷酸在培养细胞中被广泛用作基因表达的抑制剂，并已被提议作为潜在的治疗药物，但它们对其预期靶RNA的特异性程度尚不清楚。统计学考量表明，如果寡核苷酸能够通过短的或不完全互补区域在体内与mRNA分子形成杂交体，那么寡核苷酸作为反义试剂的特异性将大打折扣。我们使用非洲爪蟾卵母细胞作为模型系统来研究反义寡核苷酸在体内的潜在特异性。我们将完全匹配和部分匹配的反义寡核苷酸注射到卵母细胞中，并测量每种情况下靶RNA的降解情况。基于反义寡核苷酸在不完全匹配的靶位点导致RNA切割的程度，我们得出结论，在这个系统中，要想特异性切割预期的靶RNA而不至少部分破坏许多非靶RNA可能是不可能的。
