A Collection of Mutants for CLE-Peptide-Encoding Genes in Arabidopsis Generated by CRISPR/Cas9-Mediated Gene Targeting
A Collection of Mutants for CLE-Peptide-Encoding Genes in Arabidopsis Generated by CRISPR/Cas9-Mediated Gene Targeting
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DOI：
10.1093/pcp/pcx139
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发表时间：
2017-11
影响因子：
4.9
通讯作者：
Yasuka L. Yamaguchi;Takashi Ishida;M. Yoshimura;Yuko Imamura;Chie Shimaoka;S. Sawa
中科院分区：
文献类型：
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作者：
Yasuka L. Yamaguchi;Takashi Ishida;M. Yoshimura;Yuko Imamura;Chie Shimaoka;S. Sawa
配体-受体介导的细胞间通讯系统在多细胞生物的发育和生理活动中起着重要的协调作用。在植物中，CLAVATA 3/胚周围区（CLE）肽及其同源受体被认为参与植物生命周期的各个方面。虽然这种通信的重要性得到了广泛的认可，但大多数CLE肽的功能尚未得到表征。小信号肽编码基因的研究中的一个主要问题是由于它们的小基因大小而可获得的功能丧失突变体的数量有限。CRISPR/Cas9介导的基因靶向有可能克服这个问题，因为它可以用于在基本上任何基因中产生靶向突变，无论大小如何。在这里，我们产生了一系列的CLE-肽编码基因的突变体。新产生的clv 3和cle 40突变体分别在茎尖分生组织和根分生组织中再现了预期的突变体表型。我们的研究结果表明，CRISPR/Cas9介导的基因靶向是一种强大的遗传分析工具，即使是小基因。我们还报告了CLE 44的一种新突变体[被认为编码一种气管元件分化抑制因子（TDIF）]，并表明CLE 44有助于血管发育。这里提出的生物资源将是进一步表征CLE肽的有力工具。
The ligand-receptor-mediated intercellular communication system plays important roles in coordinating developmental and physiological events in multicellular organisms. In plants, CLAVATA3/EMBRYO SURROUNDING REGION (CLE) peptides and their cognate receptors are thought to be involved in various aspects of the plant life cycle. Although the importance of this communication is broadly recognized, most CLE peptides are yet to be functionally characterized. A major problem in research on small signaling peptide-encoding genes is the limited number of loss-of-function mutants available due to their small gene size. CRISPR/Cas9-mediated gene targeting has the potential to overcome this problem, as it can be used to generate targeted mutations in essentially any gene, regardless of size. Here we generated a series of mutants of CLE-peptide-encoding genes. Newly generated clv3 and cle40 mutants reproduced the expected mutant phenotypes in the shoot apical meristem and root meristem, respectively. Our results show that CRISPR/Cas9-mediated gene targeting is a powerful tool for genetic analyses, even of small genes. We also report a novel mutant for CLE44 [which is thought to encode a tracheary elements differentiation inhibitory factor (TDIF)] and show that CLE44 contributes to vascular development. The bioresources presented here will be a powerful tool for further characterization of CLE peptides.