Rakoczy P E
Lions Eye Institute, University of Western Australia, Nedlands, Perth, Western Australia.
Methods Mol Med. 2001;47:89-104. doi: 10.1385/1-59259-085-3:89.
Antisense DNA technology is a method to inhibit or downregulate the production of a target protein by using antisense DNA or RNA molecules. An antisense sequence is a DNA or RNA that is perfectly complementary to the target nucleotide sequence present in the cell. There are two possible mechanisms for an antisense effect. The method that relies on targeting of the mRNA is called the antisense strategy. When the double-stranded DNA or genes situated in the nucleus are targeted, the approach is called the antigene strategy. Whereas the antisense strategy is well established with several examples of in vitro and in vivo applications (1), the antigene approach is still in its infancy and our understanding of the mechanism involved is limited. The antisense strategy utilizes the ability of a 100% complementary DNA or RNA sequence to interlock or hybridize with the target mRNA thus inhibiting the translation of the target protein. This inhibition can be achieved either by blocking the binding sites for the 40S ribosomal subunit and for other translation initiation signals. Alternatively, the formation of a double-stranded DNA/RNA complex can render the RNA susceptible to RNase H digestion (2). The antigene approach is based on the binding of an antisense or sense DNA to the complimentary DNA sequence in the nucleus thus forming a triplex structure. This triplex prevents the transcription of the DNA coding sequence into mRNA (2).
反义DNA技术是一种通过使用反义DNA或RNA分子来抑制或下调靶蛋白产生的方法。反义序列是与细胞中存在的靶核苷酸序列完全互补的DNA或RNA。反义效应有两种可能的机制。依赖于靶向mRNA的方法称为反义策略。当位于细胞核中的双链DNA或基因被靶向时，这种方法称为反基因策略。虽然反义策略已经通过几个体外和体内应用的例子得到了充分确立(1)，但反基因方法仍处于起步阶段，我们对其涉及的机制的理解有限。反义策略利用100%互补的DNA或RNA序列与靶mRNA互锁或杂交的能力，从而抑制靶蛋白的翻译。这种抑制可以通过阻断40S核糖体亚基和其他翻译起始信号的结合位点来实现。或者，双链DNA/RNA复合物的形成可以使RNA易受RNase H消化(2)。反基因方法基于反义或有义DNA与细胞核中互补DNA序列的结合，从而形成三链结构。这种三链结构阻止DNA编码序列转录成mRNA(2)。
