RNA-seq技术及其在糖尿病视网膜病变中的应用

RNA-seq and its application in diabetic retinopathy

RNA-seq技术是一种使用深度测序技术的转录组学分析方法,利用高通量下一代测序技术来调查、表征和量化转录组,可以迅速全面检测某一物种特定组织特定细胞在某一状态下的几乎所有的转录本和基因序列,已广泛应用于基础研究、临床诊断以及药物研发等领域。多项研究证实其测量精度可与微阵列和定量聚合酶链式反应相媲美。RNA-seq技术可以准确检测在糖尿病视网膜病变(DR)的发病机制中发挥重要作用的mRNA和非编码RNA。利用这一技术研究DR的基因调控及分子机制,有助于寻找DR早期诊断和预测预后的生物标志物,全面系统的研究并分析特定生物学过程中的分子作用机制并寻找新的治疗靶点,对DR的基础研究和临床治疗都将具有重要意义。本文从RNA-seq技术的优势、测序平台的选择、文库制备和数据分析3个方面对该技术进行全面系统的阐述,并对该技术在DR中取得的进展进行总结和分析。

RNA-seq is a transcriptome analysis method using deep sequencing technology,which uses high-throughput next-generation sequencing technology to investigate,characterize and quantify transcriptome.It can quickly and comprehensively detect almost all transcripts and gene sequences of specific cells in specific tissue of a species,and has been widely used in basic research,clinical diagnosis and drug development and other fields.Its measurement accuracy has been confirmed comparable to microarray and quantitative polymerase chain reaction by several studies.RNA-seq technology can accurately detect mRNA and non-coding RNA which play important roles in the pathogenesis of diabetic retinopathy(DR).Using this technology to investigate the gene regulation and molecular mechanism of DR is helpful to find biomarkers for early diagnosis and prognosis of DR,to comprehensively and systematically study and analyze the molecular mechanism of specific biological process,and to find new therapeutic targets.The application of RNA-seq will be of great significance for the basic research and clinical treatment of DR.In this article,RNA-seq technology was comprehensively and systematically expounded from three aspects,which were the advantages of RNA-seq technology,the selection of sequencing platform as well as library preparation and data analysis.The progress of this technology in DR research was summarized and analyzed.