Factors influencing degradation kinetics of mRNAs and half-lives of microRNAs, circRNAs, lncRNAs in blood in vitro using quantitative PCR.
Factors influencing degradation kinetics of mRNAs and half-lives of microRNAs, circRNAs, lncRNAs in blood in vitro using quantitative PCR.
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DOI：
10.1038/s41598-022-11339-w
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发表时间：
2022-05-04
影响因子：
4.6
通讯作者：
中科院分区：
文献类型：
--
作者：
rna在样品中以及在收集、处理和测试过程中迅速降解。在本研究中，我们采用相同的方法探索了不同rna的半衰期及其影响因素，并比较了不同rna在全血和实验样品中的降解动力学和特性。新鲜抗凝血标本在室温下孵育不同时间，提取rna，并通过实时定量PCR扩增包括内参在内的基因。根据循环阈值（Ct）建立了线性半衰期模型。探讨了实验操作对提取前后RNA降解的影响。采用正交实验设计对样品和实验过程中mRNA降解的定量分析进行了探讨。血样在室温下的保存时间对RNA降解的影响最大。信使rna （mrna）的半衰期为16.4 h，环状rna （circRNAs）、长链非编码rna （lncRNAs）和微rna （miRNAs）的半衰期分别为24.56±5.2 h、17.46±3.0 h和16.42±4.2 h。RNA降解主要发生在血液样本中。mrna的半衰期是四种rna中最短的。与mrna相关的定量实验应在2 h内完成。circrna和lncrna的半衰期比前两者更长。
RNAs are rapidly degraded in samples and during collection, processing and testing. In this study, we used the same method to explore the half-lives of different RNAs and the influencing factors, and compared the degradation kinetics and characteristics of different RNAs in whole blood and experimental samples. Fresh anticoagulant blood samples were incubated at room temperature for different durations, RNAs were extracted, and genes, including internal references, were amplified by real-time quantitative PCR. A linear half-life model was established according to cycle threshold (Ct) values. The effects of experimental operations on RNA degradation before and after RNA extraction were explored. Quantitative analysis of mRNA degradation in samples and during experimental processes were explored using an orthogonal experimental design. The storage duration of blood samples at room temperature had the greatest influence on RNA degradation. The half-lives of messenger RNAs (mRNAs) was 16.4 h. The half-lives of circular RNAs (circRNAs), long non-coding RNAs (lncRNAs) and microRNAs (miRNAs) were 24.56 ± 5.2 h, 17.46 ± 3.0 h and 16.42 ± 4.2 h, respectively. RNA degradation occurred mainly in blood samples. The half-life of mRNAs was the shortest among the four kinds of RNAs. Quantitative experiments related to mRNAs should be completed within 2 h. The half-lives of circRNAs and lncRNAs were longer than those of the former two.
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影响因子：
6.8
作者：
Lippi, Giuseppe;Chance, Jeffrey J.;Simundic, Ana-Maria
通讯作者：
Simundic, Ana-Maria
影响因子：
9.3
作者：
El-Hefnawy, T;Raja, S;Godfrey, TE
通讯作者：
Godfrey, TE
影响因子：
2.7
作者：
Ju J;Huang Q;Sun J;Zhao X;Guo X;Jin Y;Ma W;Wang W
通讯作者：
Wang W
DOI：
10.1186/s12948-018-0089-4
发表时间：
2018
期刊：
Clinical and molecular allergy : CMA
影响因子：
--
作者：
Casciaro M;Di Salvo E;Brizzi T;Rodolico C;Gangemi S
通讯作者：
Gangemi S
影响因子：
9.3
作者：
McLerran, Dale;Grizzle, William E.;Semmes, O. John
通讯作者：
Semmes, O. John