Probst J C
Wilex Biotechnology GmbH, Grillparzerstrasse 10b, Munich, D-81675, Germany.
Methods. 2000 Nov;22(3):271-81. doi: 10.1006/meth.2000.1079.
The overwhelming advances of the last few years in the field of nucleic acid-based technologies laid the basis for the development of this new technology as a frontier method not only to combat diseases and infections but also to study gene function. The development of antisense strategies has generated considerable expectations in the neurosciences and, in particular, behavioral neurobiology. Antisense application in the brain has become a technology with tremendous impact, especially for determining the molecular pathways and substrates of behavior of an organism controlled by independent stimuli. The antisense agents, either oligodeoxynucleotides or ribozymes, interfere in the genetic flow of information from DNA via RNA to protein. According to the literature it seems clear that appropriately modified antisense compounds successfully and stably bind to their target ribonucleic acid molecules. This antisense binding leads to a decrease in the corresponding protein levels. If the targeted protein exerts detrimental effects on the cell or tissue, its reduction should be beneficial from a therapeutic point of view. If the investigator wants to study the function of a specific gene product the selective and transient downregulation of the corresponding target protein will help in functional analysis. In the following article I describe the chemical nature of the antisense oligodeoxynucleotides and some of the most commonly used derivatives and give some guidelines on antisense construction and application. The possible mode of action is discussed, as is expansion of the oligonucleotide-based application to ribozyme-mediated gene inhibition. Finally, problems that may be encountered during antisense application are discussed.
过去几年中，基于核酸的技术取得了巨大进展，为这项新技术的发展奠定了基础。它不仅是对抗疾病和感染的前沿方法，也是研究基因功能的前沿方法。反义策略的发展在神经科学领域，尤其是行为神经生物学领域，引发了人们相当大的期待。反义技术在大脑中的应用已成为一项具有巨大影响力的技术，特别是对于确定由独立刺激控制的生物体行为的分子途径和底物而言。反义剂，无论是寡脱氧核苷酸还是核酶，都会干扰从DNA经RNA到蛋白质的遗传信息流。根据文献，似乎很清楚的是，经过适当修饰的反义化合物能够成功且稳定地与其靶核糖核酸分子结合。这种反义结合会导致相应蛋白质水平的降低。如果靶向的蛋白质对细胞或组织产生有害影响，从治疗角度来看，其减少应该是有益的。如果研究人员想要研究特定基因产物的功能，相应靶蛋白的选择性和瞬时下调将有助于进行功能分析。在下面的文章中，我将描述反义寡脱氧核苷酸的化学性质以及一些最常用的衍生物，并给出一些关于反义构建和应用的指导方针。还将讨论可能的作用方式，以及基于寡核苷酸的应用扩展到核酶介导的基因抑制。最后，将讨论反义应用过程中可能遇到的问题。
