Lombardo A, Slack J M
Developmental Biology Programme, School of Biology and Biochemistry, University of Bath, United Kingdom.
Dev Dyn. 1997 Feb;208(2):162-9. doi: 10.1002/(SICI)1097-0177(199702)208:2<162::AID-AJA3>3.0.CO;2-G.
We studied the effects in Xenopus embryos of overexpression of antisense RNA complementary to the messenger RNA of eFGF. We show that the expression of sense RNA can be severely depressed in the presence of an excess of antisense RNA. This occurs by both partial destruction of the message and by a depression of translation of the residual message. The diminution of inducing activity of eFGF, measured in animal cap assays either by activation of the Brachyury gene or by morphology, parallels the reduction of translation. Endogenous eFGF expression is reduced to a similar extent, again by a combination of mRNA destruction and inhibition of translation. This shows that the overexpression of antisense RNA is, contrary to general opinion, a potentially useful technique for studying gene function in Xenopus embryos. However, in the case of eFGF, there is little or no overall phenotypic effect on whole embryos. This is probably because of the presence of several other FGFs with overlapping expression domains in the early embryo.
我们研究了与eFGF信使核糖核酸互补的反义核糖核酸在非洲爪蟾胚胎中的过表达效应。我们发现，在过量反义核糖核酸存在的情况下，正义核糖核酸的表达会严重受抑。这是通过部分破坏信使核糖核酸以及抑制残留信使核糖核酸的翻译来实现的。在动物帽实验中，通过激活Brachyury基因或观察形态来测量，eFGF诱导活性的降低与翻译的减少相平行。内源性eFGF的表达也以类似的程度降低，同样是通过信使核糖核酸破坏和翻译抑制的共同作用。这表明，与普遍观点相反，反义核糖核酸的过表达是研究非洲爪蟾胚胎基因功能的一种潜在有用技术。然而，就eFGF而言，对整个胚胎几乎没有或没有总体表型效应。这可能是因为在早期胚胎中存在几种其他具有重叠表达域的成纤维细胞生长因子。
