Department of Medical Biology, Faculty of Medicine, University of Szeged, Szeged, Hungary.
Department of Medical Biology, Faculty of Medicine, University of Szeged, Szeged, Hungary.
Trends Microbiol. 2019 Jul;27(7):578-592. doi: 10.1016/j.tim.2019.01.010. Epub 2019 Feb 26.
Long-read sequencing (LRS) has become increasingly popular due to its strengths in de novo assembly and in resolving complex DNA regions as well as in determining full-length RNA molecules. Two important LRS technologies have been developed during the past few years, including single-molecule, real-time sequencing by Pacific Biosciences, and nanopore sequencing by Oxford Nanopore Technologies. Although current LRS methods produce lower coverage, and are more error prone than short-read sequencing, these methods continue to be superior in identifying transcript isoforms including multispliced RNAs and transcript-length variants as well as overlapping transcripts and alternative polycistronic RNA molecules. Viruses have small, compact genomes and therefore these organisms are ideal subjects for transcriptome analysis with the relatively low-throughput LRS techniques. Recent LRS studies have multiplied the number of previously known transcripts and have revealed complex networks of transcriptional overlaps in the examined viruses.
长读测序(LRS)因其在从头组装和解决复杂 DNA 区域以及确定全长 RNA 分子方面的优势而变得越来越流行。在过去几年中,已经开发出两种重要的 LRS 技术,包括 Pacific Biosciences 的单分子实时测序和 Oxford Nanopore Technologies 的纳米孔测序。尽管当前的 LRS 方法的覆盖度较低,并且比短读测序更容易出错,但这些方法在识别转录异构体方面仍然具有优势,包括多拼接 RNA 和转录长度变体以及重叠转录物和多顺反子 RNA 分子。病毒具有小而紧凑的基因组,因此这些生物体是相对低通量 LRS 技术进行转录组分析的理想对象。最近的 LRS 研究增加了以前已知的转录本数量,并揭示了所研究病毒中复杂的转录重叠网络。