NEIBank: Est Analysis & Bioinformatics For Ocular Genomi

眼睛是一个复杂的系统，由各种发育起源的高度分化的组织组成。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。大多数人类表达基因只能通过表达序列标签（est）来了解。由于眼部组织在cDNA文库中的代表性较差，因此开始了一个名为NEIBank的项目，以提高EST的覆盖范围并开发眼部分子百科全书。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，并通过深度测序进行了检测。使用定制软件GRIST对每个非规范化库中的2000到10000个克隆进行了测序、分析和聚类。建立了几个重要动物模型库，包括大鼠的角度、视网膜和全眼，小鼠的全眼、泪腺、视网膜和RPE，狗和兔的前后节，斑马鱼的晶状体和全眼。许多新的基因和遗传变异已经被发现。在一个包含关键字和染色体定位数据的基于web的数据库（http://neibank.nei.nih.gov）中，总共对大约40,000个眼部cDNA克隆进行了测序和组织。这项工作产生的人类DNA资源已被用于创建用于眼部研究的微阵列。一个含有超过13,000个非冗余人眼表达序列的cDNA微阵列已经构建并正在测试中。

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.