Clarke M L, Patrikakis M, Atkins D
Johnson and Johnson Research Pty Limited, Locked Bag 4555, Strawberry Hill, NSW, 2012, Australia.
Biochem Biophys Res Commun. 2000 Feb 5;268(1):8-13. doi: 10.1006/bbrc.2000.2085.
The fission yeast Schizosaccharomyces pombe has recently been established as an experimental model for the study of antisense RNA-mediated gene suppression. To validate the use of S. pombe as a host for identifying antisense genes for use in human cells, it was important to determine if sequences identified in yeast were as equally effective in a human cell line. This report describes the comparison of a range of lacZ antisense RNAs targeting a lacZ gene expressed in HeLa cells in a comparable manner to its expression in S. pombe cells in earlier studies. In both cell types, the same lacZ gene target was expressed using the same promoter. Antisense genes were expressed episomally in both experimental systems and the levels of suppression determined. In all cases, the relative level of suppression of the lacZ gene was similar in the mammalian and yeast cells. This result indicates that, at least for lacZ antisense RNA, results obtained in fission yeast are predictive of their behavior in the mammalian cellular environment.
粟酒裂殖酵母最近已被确立为研究反义RNA介导的基因抑制的实验模型。为了验证粟酒裂殖酵母作为宿主用于鉴定人类细胞中反义基因的用途，确定酵母中鉴定出的序列在人类细胞系中是否同样有效很重要。本报告描述了一系列针对在HeLa细胞中表达的lacZ基因的lacZ反义RNA的比较，其表达方式与早期研究中在粟酒裂殖酵母细胞中的表达方式相当。在两种细胞类型中，使用相同的启动子表达相同的lacZ基因靶标。反义基因在两个实验系统中以附加体形式表达，并确定抑制水平。在所有情况下，lacZ基因的相对抑制水平在哺乳动物细胞和酵母细胞中相似。这一结果表明，至少对于lacZ反义RNA而言，在裂殖酵母中获得的结果可预测其在哺乳动物细胞环境中的行为。
