Maltese J Y, Sharma H W, Vassilev L, Narayanan R
Division of Oncology, Roche Research Center, Hoffmann-La Roche Inc., Nutley, NJ 07110, USA.
Nucleic Acids Res. 1995 Apr 11;23(7):1146-51. doi: 10.1093/nar/23.7.1146.
The use of antisense oligomers to achieve inhibition of gene expression is complicated by frequent non-specific effects, and even the control oligomers often exhibit sequence-specific effects. We have recently shown that in diverse tumor-derived cell lines, a 24mer phosphorothioate oligomer antisense to the relA subunit of NF-kappa B transcription factor causes a block of cellular adhesion, inhibition of nuclear NF-kappa B and Sp1 DNA-binding activity and inhibition of tumor cell growth in vitro and in vivo. In this study we use the same model to attempt to define the limits of antisense specificity. We demonstrate that single base pair substitution can virtually abolish the antisense activity. The relative position of mismatches within the antisense sequence is critical to the loss of activity. Our results further indicate that antisense specificity is determined not only by the content of the sequence but also by its occurrence with reference to the surrounding sequences.
使用反义寡聚物来实现基因表达抑制会因频繁的非特异性效应而变得复杂，甚至对照寡聚物也常常表现出序列特异性效应。我们最近发现，在多种肿瘤来源的细胞系中，一种针对NF-κB转录因子RelA亚基的24聚体硫代磷酸酯寡聚物反义物会导致细胞黏附受阻、核NF-κB和Sp1 DNA结合活性受到抑制，以及在体外和体内抑制肿瘤细胞生长。在本研究中，我们使用相同的模型来试图确定反义特异性的限度。我们证明单碱基对替换实际上可以消除反义活性。反义序列内错配的相对位置对于活性丧失至关重要。我们的结果进一步表明，反义特异性不仅由序列内容决定，还由其相对于周围序列的出现情况决定。
