NEIBank: Est Analysis & Bioinformatics For Ocular Genomi

NEIBank：Est 分析

• 批准号: 6826739
• 负责人: Graeme J Wistow
• 金额: --
• 依托单位: NATIONAL EYE INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6826739
• 关键词: animal genetic material tag; animal tissue; complementary DNA; computer assisted sequence analysis; computer program /software; cornea; eye; eye disorder; gene expression; genetic library; genetic mapping; genetic registry /resource /referral center; genetically modified animals; genome; human genetic material tag; human tissue; iris; laboratory mouse; microarray technology; molecular biology information system; retina; sequence tagged sites

The eye is a complex system of highly differentiated tissues of various developmental origins. Many genes essential for eye function are tissue-specific and many of those known are associated with genetic eye diseases. The majority of human expressed genes are known only through expressed sequence tags (ESTs). Since eye tissues were poorly represented in the cDNA libraries initially contributed to dbEST a project called NEIBank was begun to improve the EST coverage and to develop a molecular encyclopedia for the eye. cDNA libraries for several human eye tissues, including lens, iris, RPE/choroid, retina, trabecular meshwork, lacrimal gland and keratoconus cornea have been constructed and examined by in-depth sequencing. Between 2,000 and 10,000 clones from each un-normalized library have been sequenced, analyzed and clustered using custom software, GRIST. Several libraries have been made for important animal models, including angle, retina and whole eye of rat, whole eye, lacrimal gland, retina and RPE of mouse, anterior and posterior segments for dog and rabbit and lens and whole eye for zebrafish . Many novel genes and genetic variants have been identified. In total, approximately 40,000 ocular cDNA clones have been sequenced and organized in a web-based database (http://neibank.nei.nih.gov) that contains keyword and chromosomal location data. The human DNA resource generated by this work have been used to create micro-arrays for ocular studies. A cDNA microarray with over 13,000 non-redundant human eye expressed sequences has been constructed and is being tested. Several novel genes discovered through NEIBank have been selected for verification and further study. These include lengsin an abundant novel lens specific transcript related to glutamine synthetase and iris-expressed growth factor. Lengsin expression is associated with terminal differentiation and nuclear breakdown in the lens. Recombinant mouse and human proteins are being used for function and structure studies and transgenic mice using the lengsin promoter have been made. A knock-mouse model for lengsin is under construction using "recombineering" technology. We have shown that IEGF/PDGFD has a key role in control of lens growth and development of the anterior segment. Antibody to IEGF blocks lens cell proliferation and may have translational research applications in secondary and subcapsular cataracts. This factor is also specifically expressed in the retina and may have a role in photoreceptor health. Analysis of a new keratoconus cornea library has shown that the library contains an excellent representation of human cornea expressed genes but has also revealed an important candidate gene for involvement in this condition. The leads from these data are being pursued, along with several other novel gene products including retbindin and Mp19ins. Retbindin may be the carotenoid binding protein of the retina, while Mp19ins may have a specific role in correct formation of the human lens.

眼睛是一个由不同发育来源的高度分化的组织组成的复杂系统。许多对眼睛功能至关重要的基因是组织特异性的，其中许多已知的基因与遗传性眼病有关。大多数人类表达的基因都是通过表达序列标签(EST)才知道的。由于眼部组织最初在cDNA文库中的代表性很差，因此开始了一项名为NEIBank的项目，以提高EST的复盖率，并为眼睛开发一部分子百科全书。我们已经构建了包括晶状体、虹膜、RPE/脉络膜、视网膜、小梁网、泪腺和圆锥角膜在内的几种人眼组织的cDNA文库，并进行了深度测序。来自每个非标准化文库的2,000到10,000个克隆已经使用定制软件GRIST进行了测序、分析和聚类。建立了大鼠的角度、视网膜和全眼，小鼠的全眼、泪腺、视网膜和RPE，狗和兔的前节和后节，斑马鱼的晶状体和全眼等重要动物模型文库。许多新的基因和遗传变异已经被发现。总共，大约40,000个眼部cDNA克隆被测序并组织在一个基于网络的数据库(http://neibank.nei.nih.gov))中，该数据库包含关键字和染色体位置数据。这项工作产生的人类DNA资源已被用于创建眼部研究的微阵列。一个包含13,000多个非冗余人眼表达序列的基因芯片已经构建完成，并正在进行测试。 通过NEIBank发现的几个新基因已经被选中进行验证和进一步研究。这些包括Lengin，一个丰富的与谷氨酰胺合成酶和虹膜表达的生长因子相关的新的晶状体特异性转录本。Lengsin的表达与晶状体的终末分化和核破裂有关。重组鼠蛋白和人蛋白正被用于功能和结构研究，并已获得使用长度蛋白启动子的转基因小鼠。利用“重组工程”技术，正在建造一种长度蛋白的敲打老鼠模型。我们已经证明IEGF/PDGFD在控制晶状体的生长和前段的发育中起着关键作用。IEGF抗体可阻断晶状体细胞的增殖，并可能在继发性白内障和包膜下白内障的翻译研究中应用。这一因子也在视网膜中特异表达，并可能在感光器健康中发挥作用。对一个新的圆锥角膜文库的分析表明，该文库包含了良好的人类角膜表达基因，但也揭示了与这种情况有关的一个重要候选基因。来自这些数据的线索正在与其他几种新的基因产品一起寻找，包括视黄素和Mp19ins。Retbindin可能是视网膜的类胡萝卜素结合蛋白，而Mp19可能在人类晶状体的正确形成中具有特定的作用。