Crop Bioinformatics, Institute of Crop Science and Resource Conservation (INRES), University of Bonn, 53115 Bonn, Germany.
Crop Functional Genomics, Institute of Crop Science and Resource Conservation (INRES), University of Bonn, 53115 Bonn, Germany.
Bioinformatics. 2022 Jan 12;38(3):837-838. doi: 10.1093/bioinformatics/btab679.
Insertional mutagenesis allows for the creation of loss-of-function mutations on a genome-wide scale. In theory, every gene can be 'knocked out' via the insertion of an additional DNA sequence. Resources of sequence-indexed mutants of plant and animal model organisms are instrumental for functional genomics studies. Such repositories significantly speed up the acquisition of interesting genotypes and allow for the validation of hypotheses regarding phenotypic consequences in reverse genetics. To create such resources, comprehensive sequencing of flanking sequence tags using protocols such as Mutant-seq requires various downstream computational tasks, and these need to be performed in an efficient and reproducible manner.
Here, we present MuWU, an automated Mutant-seq workflow utility initially created for the identification of Mutator insertion sites of the BonnMu resource, representing a reverse genetics mutant collection for functional genetics in maize (Zea mays). MuWU functions as a fast, one-stop downstream processing pipeline of Mutant-seq reads. It takes care of all complex bioinformatic tasks, such as identifying tagged genes and differentiating between germinal and somatic mutations/insertions. Furthermore, MuWU automatically assigns insertions to the corresponding mutated seed stocks. We discuss the implementation and how parameters can easily be adapted to use MuWU for other species/transposable elements.
MuWU is a Snakemake-based workflow and freely available at https://github.com/tgstoecker/MuWU.
Supplementary data are available at Bioinformatics online.
插入突变可在全基因组范围内创建功能丧失突变。从理论上讲,每个基因都可以通过插入额外的 DNA 序列“敲除”。植物和动物模式生物的序列索引突变体资源对于功能基因组学研究至关重要。这些资源大大加快了获得有趣基因型的速度,并允许在反向遗传学中验证关于表型后果的假设。为了创建这些资源,需要使用 Mutant-seq 等协议对侧翼序列标签进行全面测序,这需要执行各种下游计算任务,并且需要以高效和可重复的方式执行。
在这里,我们介绍了 MuWU,这是一种自动化的 Mutant-seq 工作流程实用程序,最初是为识别 BonnMu 资源的 Mutator 插入位点而创建的,该资源代表了用于玉米(Zea mays)功能遗传学的反向遗传学突变体集合。MuWU 作为 Mutant-seq 读取的快速一站式下游处理管道。它负责所有复杂的生物信息学任务,例如识别标记基因和区分生殖突变和体细胞突变/插入。此外,MuWU 自动将插入分配给相应的突变种子库存。我们讨论了实现方式以及如何轻松调整参数,以便将 MuWU 用于其他物种/转座元件。
MuWU 是一个基于 Snakemake 的工作流程,可在 https://github.com/tgstoecker/MuWU 上免费获得。
补充数据可在 Bioinformatics 在线获得。