Tamilselvan-Nattar-Amutha Suriya, Hiekel Stefan, Hartmann Franziska, Lorenz Jana, Dabhi Riddhi Vijay, Dreissig Steven, Hensel Goetz, Kumlehn Jochen, Heckmann Stefan
Leibniz Institute of Plant Genetics and Crop Plant Research (IPK) OT Gatersleben, Seeland, Germany.
Front Plant Sci. 2023 Jun 19;14:1201446. doi: 10.3389/fpls.2023.1201446. eCollection 2023.
Genome editing strategies in barley () typically rely on -mediated genetic transformation for the delivery of required genetic reagents involving tissue culture techniques. These approaches are genotype-dependent, time-consuming, and labor-intensive, which hampers rapid genome editing in barley. More recently, plant RNA viruses have been engineered to transiently express short guide RNAs facilitating CRISPR/Cas9-based targeted genome editing in plants that constitutively express . Here, we explored virus-induced genome editing (VIGE) based on barley stripe mosaic virus (BSMV) in -transgenic barley. Somatic and heritable editing in the gene () resulting in albino/variegated chloroplast-defective barley mutants is shown. In addition, somatic editing in meiosis-related candidate genes in barley encoding ASY1 (an axis-localized HORMA domain protein), MUS81 (a DNA structure-selective endonuclease), and ZYP1 (a transverse filament protein of the synaptonemal complex) was achieved. Hence, the presented VIGE approach using BSMV enables rapid somatic and also heritable targeted gene editing in barley.
大麦（）中的基因组编辑策略通常依赖于介导的遗传转化来递送所需的遗传试剂，这涉及组织培养技术。这些方法依赖基因型、耗时且 labor-intensive，这阻碍了大麦中的快速基因组编辑。最近，植物RNA病毒已被改造以瞬时表达短引导RNA，从而促进在组成型表达的植物中基于CRISPR/Cas9的靶向基因组编辑。在这里，我们在转基因大麦中探索了基于大麦条纹花叶病毒（BSMV）的病毒诱导基因组编辑（VIGE）。结果显示，基因（）中的体细胞编辑和可遗传编辑导致了白化/杂色叶绿体缺陷型大麦突变体。此外，在大麦中编码ASY1（一种位于轴上的HORMA结构域蛋白）、MUS81（一种DNA结构选择性核酸内切酶）和ZYP1（联会复合体的横向细丝蛋白）的减数分裂相关候选基因中实现了体细胞编辑。因此，所展示的使用BSMV的VIGE方法能够在大麦中实现快速的体细胞和可遗传靶向基因编辑。
