Identification and Quantification of Small RNAs.
Identification and Quantification of Small RNAs.
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DOI：
10.1007/978-1-0716-0880-7_11
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Zhang X
中科院分区：
文献类型：
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作者：
Sun D;Ma Z;Zhu J;Zhang X
RNA沉默在植物的生长、发育以及对非生物和生物胁迫的反应等多种生物过程中起着关键作用。RNA沉默是由长度为21-24个核苷酸（nt）的小非编码RNA （sRNAs）引导，通过转录或转录后基因沉默机制，将其装载到Argonaute （AGO）中，抑制靶位点和转录物的表达。sRNAs的鉴定和定量表征是了解其生物学功能的关键步骤。在这里，我们制定了一个循序渐进的协议，以精确地说明sRNA文库的克隆过程和相应的计算分析恢复的sRNA。该方案可用于包括拟南芥在内的各种生物，并与各种高通量序列技术（如Illumina Hiseq）兼容。因此，我们希望该方案代表了一种准确的方法来鉴定和量化体内的srna。
RNA silencing plays a critical role in diverse biological processes in plants including growth, development, and responses to abiotic and biotic stresses. RNA silencing is guided by small non-coding RNAs (sRNAs) with the length of 21–24 nucleotides (nt) that are loaded into Argonaute (AGO) to repress expression of target loci and transcripts through transcriptional or posttranscriptional gene silencing mechanisms. Identification and quantitative characterization of sRNAs are crucial steps toward appreciation of their functions in biology. Here, we developed a step-by-step protocol to precisely illustrate the process of cloning of sRNA libraries and correspondingly computational analysis of the recovered sRNAs. This protocol can be used in all kinds of organisms, including Arabidopsis, and is compatible with various high-throughput sequence technologies such as Illumina Hiseq. Thus, we wish that this protocol represents an accurate way to identify and quantify sRNAs in vivo.