Dunn Simon R, Phillips Wendy S, Green Douglas R, Weis Virginia M
Department of Zoology, Oregon State University, Corvallis, Oregon 97331, USA.
Biol Bull. 2007 Jun;212(3):250-8. doi: 10.2307/25066607.
Since the discovery of the ancient eukaryotic process of RNA-mediated gene silencing, the reverse-genetics technique RNA interference (RNAi) has increasingly been used to examine gene function in vertebrate and invertebrate systems. In this study, we report on the use of RNAi, adapted from studies on animal model systems, to manipulate gene expression in a symbiotic marine cnidarian. We describe gene knockdown of actin and of acasp--a cysteine protease, or caspase--in the symbiotic sea anemone Aiptasia pallida. Knockdown was assessed qualitatively with in situ hybridizations for both genes. Quantitative PCR and caspase activity assays were used as a quantitative measure of knockdown for acasp.
自从发现古老的RNA介导的基因沉默真核生物过程以来,反向遗传学技术RNA干扰(RNAi)越来越多地用于研究脊椎动物和无脊椎动物系统中的基因功能。在本研究中,我们报告了借鉴动物模型系统研究而采用的RNAi技术,用于操纵共生海洋刺胞动物中的基因表达。我们描述了在共生海葵苍白艾氏海葵中肌动蛋白和一种半胱氨酸蛋白酶(即胱天蛋白酶)acasp的基因敲低情况。通过对这两个基因进行原位杂交来定性评估基因敲低情况。使用定量PCR和胱天蛋白酶活性测定作为acasp基因敲低的定量指标。