Comprehensive analysis of 300 KAO-NASHI (Face less) genes.

人类基因组计划提供了人类基因组中23000个蛋白质编码基因的计算机估计。然而，许多这些蛋白质编码基因的存在并没有得到实验证据的充分证明。一般来说，具有已知基序的蛋白质很容易分类，但大量蛋白质在其序列中没有明显的基序。我们将这些没有明显基序的基因/蛋白命名为kao - nashi (face -less)，并启动了一个项目，通过比较基因组学和敲除分析来揭开它们的脸（花脸）。通过InterPro基序分析、BLAST同源性检索和PubMed文献检索，从人类基因组序列中提取了1000个KAO-NASHI基因。选择一种小鱼（Oryzias latipes）作为实验系统，敲除具有morpholino-反义寡核苷酸的人类KAO-NASHI基因的medaka同源物。作为初步的研究，我们设计了反义寡核苷酸来靶向130个日本稻花菜基因的翻译起始位点。将这些反义寡核苷酸微量注入水母胚胎后，会干扰其早期发育阶段的形态发生，并观察到一定速率的形态变化。因此，我们获得了这些medaka kao-nashi基因如何参与胚胎模式和器官形成过程的初步粗略信息。迄今为止，已经发现了60个导致胚胎缺陷的基因，这些基因根据发育亚阶段和表达谱进一步分类。因此，我们使用medaka的方法似乎是有效的，它最终将提供关于人类KAO-NASHI基因/蛋白质的功能信息。

The human genome project has provided a computer-estimation of 23,000 protein-coding genes in the human genome. However, many of these protein-coding genes are not fully proven for their existence by experimental evidence. In general, proteins with known motifs are readily classified, but substantial numbers of protein have no obvious motifs in their sequences. We designated these genes/proteins without obvious motifs as KAO-NASHI (Face-less) and initiated a project to unveil their face (kao) by comparative genomics and knock-down analysis.We extracted 1,000 KAO-NASHI genes from human genome sequence by step-wise filtration with InterPro motif analysis, BLAST homology search and PubMed document search. A small fish medaka (Oryzias latipes) was chosen as an experimental system to knock-down medaka orthologs of human KAO-NASHI genes with morpholino-antisense oligos. As an initial study, we designed antisense oligos to target translation initiation sites of 130 medaka kao-nashi genes. When these antisense oligos were micro-injected into medaka embryos, their morphogenesis at early developmental stages was disturbed and morphological changes were observed at certain rates. Thus, we obtained initial sketchy information how these medaka kao-nashi genes are involved in the embryonic process of patterning and organ formation. To date, 60 genes were found to cause embryonic defects and these were further classified in terms of developmental sub-stages and expression profiles. Thus, our approach using medaka seems effective and it will eventually provide functional information on the human KAO-NASHI genes/proteins.