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表达:一个涵盖眼部组织多个发育阶段已知和新转录本的转录组谱数据库。

Express: A database of transcriptome profiles encompassing known and novel transcripts across multiple development stages in eye tissues.

机构信息

Department of BioHealth Informatics, School of Informatics and Computing, Indiana University Purdue University, 719 Indiana Ave Ste 319, Walker Plaza Building, Indianapolis, IN 46202, United States.

Department of Biological Sciences, University of Delaware, Newark, DE 19716, United States.

出版信息

Exp Eye Res. 2018 Mar;168:57-68. doi: 10.1016/j.exer.2018.01.009. Epub 2018 Jan 11.

Abstract

Advances in sequencing have facilitated nucleotide-resolution genome-wide transcriptomic profiles across multiple mouse eye tissues. However, these RNA sequencing (RNA-seq) based eye developmental transcriptomes are not organized for easy public access, making any further analysis challenging. Here, we present a new database "Express" (http://www.iupui.edu/∼sysbio/express/) that unifies various mouse lens and retina RNA-seq data and provides user-friendly visualization of the transcriptome to facilitate gene discovery in the eye. We obtained RNA-seq data encompassing 7 developmental stages of lens in addition to that on isolated lens epithelial and fibers, as well as on 11 developmental stages of retina/isolated retinal rod photoreceptor cells from publicly available wild-type mouse datasets. These datasets were pre-processed, aligned, quantified and normalized for expression levels of known and novel transcripts using a unified expression quantification framework. Express provides heatmap and browser view allowing easy navigation of the genomic organization of transcripts or gene loci. Further, it allows users to search candidate genes and export both the visualizations and the embedded data to facilitate downstream analysis. We identified total of >81,000 transcripts in the lens and >178,000 transcripts in the retina across all the included developmental stages. This analysis revealed that a significant number of the retina-expressed transcripts are novel. Expression of several transcripts in the lens and retina across multiple developmental stages was independently validated by RT-qPCR for established genes such as Pax6 and Lhx2 as well as for new candidates such as Elavl4, Rbm5, Pabpc1, Tia1 and Tubb2b. Thus, Express serves as an effective portal for analyzing pruned RNA-seq expression datasets presently collected for the lens and retina. It will allow a wild-type context for the detailed analysis of targeted gene-knockout mouse ocular defect models and facilitate the prioritization of candidate genes from Exome-seq data of eye disease patients.

摘要

测序技术的进步促进了多个小鼠眼组织中核苷酸分辨率的全基因组转录组谱的研究。然而,这些基于 RNA 测序(RNA-seq)的眼发育转录组并未进行组织整理,不利于公众访问,使得进一步的分析变得具有挑战性。在这里,我们提出了一个新的数据库“Express”(http://www.iupui.edu/∼sysbio/express/),它统一了各种小鼠晶状体和视网膜的 RNA-seq 数据,并提供了转录组的用户友好可视化,以促进眼内基因的发现。我们从公开的野生型小鼠数据集获得了涵盖晶状体 7 个发育阶段的 RNA-seq 数据,以及分离的晶状体上皮和纤维以及 11 个视网膜/分离的视网膜杆状光感受器细胞发育阶段的 RNA-seq 数据。这些数据集经过预处理、比对、定量和表达水平标准化,以识别已知和新的转录本。Express 提供了热图和浏览器视图,方便转录本或基因座的基因组组织导航。此外,它允许用户搜索候选基因,并将可视化和嵌入数据导出,以方便下游分析。我们在所有包括的发育阶段中总共鉴定出了 >81,000 个在晶状体中的转录本和 >178,000 个在视网膜中的转录本。该分析表明,相当数量的视网膜表达转录本是新的。在多个发育阶段,通过 RT-qPCR 独立验证了 Pax6 和 Lhx2 等已建立基因以及 Elavl4、Rbm5、Pabpc1、Tia1 和 Tubb2b 等新候选基因在晶状体和视网膜中的表达。因此,Express 作为一个有效的门户,可用于分析目前为晶状体和视网膜收集的修剪 RNA-seq 表达数据集。它将为靶向基因敲除小鼠眼缺陷模型的详细分析提供野生型背景,并有助于从眼疾病患者的外显子组测序数据中优先考虑候选基因。

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