在本项研究中，作者仅使用了2.5 μg 的骨髓瘤临床样品total RNA（direct RNA-seq 推荐量：50 μg）用于xPore m6A位点检测，利用METTL3-KO HEK293T细胞作为未修饰对照进行xPore模型建立，克服了临床样品缺乏相应对照组的问题。综上，该研究开发了一种基于Nanopore direct RNA测序的RNA甲基化差异分析计算方法，实现...
1. 什么是Nanopore 测序 测序大家都并不陌生,通过RNA-seq等测序技术我们了解了很多调控的信息。但是这些测序往往是短读长(short read)或者是cDNA测序的结果,对于一些链特异性的结果,我们需要通过对应的测序技术等等。nanopore是第三代的测序技术,主要通过在nanopore周围添加电解液,电压驱动以及驱动蛋白牵引带电粒子的方式...
参考文献: [1]Parker M T , Knop K , Sherwood A V , et al. Nanopore direct RNA sequencing maps the complexity of Arabidopsis mRNA processing and m6A modification[J]. Elife Sciences, 2020, 9:e49658. [2]Froussios K , Kira Mourão, Simpson G , et al. Relative Abundance of Trans ( ...
Nanopore is the only technology that can directly sequence RNA strands without reverse transcription or PCR reactions. Nanopore direct RNA sequencing technology sequences natural RNA, allowing it to retain and detect RNA base modification information, estimate the poly(A) tail length relatively accurately...
为了将RNA直接测序应用于单位点分辨率的RNA修饰从头测序,研究员研发了利用纳米孔RNA直接测序的m6A鉴定软件(MINES),对HEK293T细胞转录本进行了分析,鉴定超过13,000个之前未注释的DRACH位点的m6A甲基化状态;还在人乳腺上皮细胞(包括异构体)中鉴定得40,000个位点,这些位点分别对m6A writer、METTL3、eraser、ALKBH5敏感。
本周一,Nature Methods在线发表Oxford Nanopore direct RNA测序技术测评文章,结果表明,【不经反转、无须扩增的RNA直接测序】能获得全长的链特异性RNA,无测序偏好性,并同时记录碱基修饰,为后续研究基因结构和基因表达,提供新技术新方法。 direct RNA建库测序
该研究应用Nanopore Direct RNA测序技术研究中华鳖转录组的复杂性。与M表型相比,F表型可变剪接差异显著。m5C和m6A两种RNA甲基化修饰在不同性别表型之间存在差异,并对关键基因的m6A甲基化水平进行了验证。该研究揭示了生物表观遗传在中华鳖性逆转过程中扮演的重要作用。
近日,福建农林大学海峡联合研究院林学中心顾连峰教授研究组在Plant Physiology发表了题为“Drought induces epitranscriptome and proteome changes in stem-differentiating xylem of Populus trichocarpa”的研究论文,揭示了毛果杨次生木质部中RNA全长转录本比率、N6-甲基腺嘌呤(N6-methyladenosine, m6A)修饰率、Poly(A)位点...
Standard direct RNA-sequencing (RNA-seq) relies on poly(A) selection by using a poly(T)-tethering adaptor that base-pairs canonically to a motor protein linked adaptor, facilitating the movement of the polynucleotide string through the protein pore (Fig.1A). While standard direct RNA-seq provid...
Characterizing complex viral transcriptomes by conventional RNA sequencing approaches is complicated by high gene density, overlapping reading frames, and complex splicing patterns. Direct RNA sequencing (direct RNA-seq) using nanopore arrays offers an e