转录组测序中核糖体RNA消减策略：研究进展与方法比较

doi:10.16431/j.cnki.1671-7236.2025.11.002

摘要/Abstract

摘要： 转录组测序(RNA sequencing，RNA-Seq)技术是一种能从细胞内基因和调控元件的转录水平揭示各种复杂生物学现象的有力工具，在农业育种、品种资源、营养代谢及医学等领域的应用十分广泛。核糖体RNA(rRNA)占细胞总RNA的80%以上，若直接对样品进行RNA-Seq建库测序，获得的测序数据超过90%来自rRNA，因此很难检测到各类低丰度RNA。rRNA消减技术能大大提高RNA-Seq技术的基因检测效率和敏感性，并能对低丰度非编码RNA进行定量检测，为揭示一些复杂的生物学机制提供更为精准的数据支持，而该技术在RNA-Seq建库中的作用常常没有得到应有的重视，导致部分测序数据不能反映样品的真实值。笔者对RNA-Seq中几种rRNA消减策略进行了概述，包括Olig dT磁珠捕获法、“拉出”法、RNase H选择性消减法和耐热双特异性核酸酶(DSN)选择性消减法，以及非随机引物反转录法、RNA阻断引物法等，并对各自的优劣势进行了比较，旨在为采用RNA-Seq技术的研究人员选择最合适的rRNA消减策略提供参考，为获得最接近样品真实值的研究数据提供支持和参考，以更好地服务于科学研究和技术创新。

关键词: 转录组测序; 核糖体RNA; 消减策略

Abstract: RNA sequencing (RNA-Seq) technology is a powerful tool that reveals various complex biological phenomena by analyzing the transcriptional levels of genes and regulatory elements within cells.It has a wide range of applications in fields such as agricultural breeding,variety resources,nutrition metabolism and medicine.Ribosomal RNA (rRNA) accounts for over 80% of the total RNA in cells. Direct RNA-Seq library sequencing of samples yields data in which more than 90% of the sequences from rRNA,making it difficult to detect various low abundance RNAs.rRNA reduction technology can significantly improve the gene detection efficiency and sensitivity of RNA-Seq technology,and can quantitatively detect low abundance non-coding RNA,providing more accurate data support for revealing some complex biological mechanisms.However,the role of this technology in RNA-Seq library construction has often not been given due attention,resulting in some sequencing data not reflecting the true value of the sample.The authors provide an overview of several rRNA reduction strategies in RNA-Seq,including Olig dT magnetic bead capture method,“pull-out” method,RNase H selective reduction method,and thermostable duplex-specific nuclease (DSN) selective method, non random primer reverse transcription method,and RNA blocking primer methods,etc.The advantages and disadvantages of each strategy are compared,and intended to select the most suitable ribosomal RNA reduction strategy for researchers employing RNA-Seq technology,provide support and reference for obtaining research data that is closest to the true value of the sample,and enable new technologies to better serve scientific research and technological innovation.

Key words: RNA-Seq; rRNA; strategies reduction

中图分类号:

S961.6

李慧彤, 刘丹, 高建帅, 唐新月, 张博源, 蒋卉, 范学政, 张广智, 丁家波, 沈青春, 熊涛. 转录组测序中核糖体RNA消减策略：研究进展与方法比较[J]. 中国畜牧兽医, 2025, 52(11): 5030-5039.

LI Huitong, LIU Dan, GAO Jianshuai, TANG Xinyue, ZHANG Boyuan, JIANG Hui, FAN Xuezheng, ZHANG Guangzhi, DING Jiabo, SHEN Qingchun, XIONG Tao. Ribosomal RNA Reduction Strategies in RNA-Seq: Research Progress and Method Comparison[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(11): 5030-5039.

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