Knyazev Andrey, Glushkevich Anna, Fesenko Igor
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, 16/10, Ulitsa Miklukho-Maklaya, Moscow, 117997, Russian Federation.
Data Brief. 2020 Nov 29;33:106602. doi: 10.1016/j.dib.2020.106602. eCollection 2020 Dec.
Nonsense-mediated mRNA decay (NMD) is a system that controls the quality of mRNA transcripts in eukaryotes by degradation of aberrant transcripts in a pioneer round of translation. In mammals, NMD targets one-third of mutated, disease-causing mRNAs and ∼10% of unmutated mRNAs, facilitating appropriate cellular responses to environmental changes [1]. In plants, NMD plays an important role in development and regulating abiotic and biotic stress responses [2]. The transcripts with premature termination codons (PTCs), upstream ORFs or long 3'-UTRs can be targeted to NMD. It was shown that alternative splicing plays a crucial role in regulation of NMD triggering, for example, by the introduction of a PTC in transcripts. Therefore, the correct identification of mRNA isoforms is a key step in the study of the principles of regulation of the cell transcriptome by the NMD pathway. Here, we performed long-read sequencing of Physcomitrella () mutant Δ line 2 native transcriptome by Oxford Nanopore Technology (ONT). The Δ is a knockout (KO) mutant deficient in SMG1 kinase is a key component of NMD system in plants and animals [3]. RNA was isolated with Trizol from 5 day old protonemata and sequenced using kit SQK-RNA002, flow cells FLO-MIN106 and a MinION device (Oxford Nanopore Technologies Ltd., UK (ONT)) in three biological repeats. Basecalling was performed with Guppy v.4.0.15. The presented transcriptomes give advantages in the identification and functional characterization of RNA transcripts that are direct targets of the Nonsense-mediated mRNA decay system.
无义介导的mRNA降解(NMD)是一种通过在翻译的起始轮中降解异常转录本来控制真核生物中mRNA转录本质量的系统。在哺乳动物中,NMD靶向三分之一的突变致病mRNA和约10%的未突变mRNA,促进细胞对环境变化做出适当反应[1]。在植物中,NMD在发育以及调节非生物和生物胁迫反应中发挥重要作用[2]。带有提前终止密码子(PTC)、上游开放阅读框(ORF)或长3'非翻译区(UTR)的转录本可被NMD靶向。研究表明,可变剪接在NMD触发的调控中起关键作用,例如,通过在转录本中引入PTC。因此,正确识别mRNA异构体是研究NMD途径调控细胞转录组原理的关键步骤。在此,我们通过牛津纳米孔技术(ONT)对小立碗藓()突变体Δ株系2的天然转录组进行了长读长测序。Δ是一种敲除(KO)突变体,缺乏SMG1激酶,而SMG1激酶是动植物NMD系统的关键组成部分[3]。使用Trizol从5日龄的原丝体中分离RNA,并使用试剂盒SQK-RNA002、流动槽FLO-MIN106和MinION设备(英国牛津纳米孔技术有限公司(ONT))进行三次生物学重复测序。使用Guppy v.4.0.15进行碱基识别。所呈现的转录组在识别和功能表征作为无义介导的mRNA降解系统直接靶标的RNA转录本方面具有优势。