CRISPR/Cas9-mediated mutagenesis of lncRNA1459 alters tomato fruit ripening
CRISPR/Cas9-mediated mutagenesis of lncRNA1459 alters tomato fruit ripening
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CRISPR/Cas9介导的lncRNA1459诱变改变番茄果实成熟
DOI：
10.1111/tpj.13872
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发表时间：
2018-05-01
期刊：
影响因子：
7.2
通讯作者：
Zhu, Hongliang
中科院分区：
文献类型：
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作者：
Li, Ran;Fu, Daqi;Zhu, Hongliang
随着高通量测序技术的发展，许多长链非编码RNA（lncRNA）被发现在多种生物过程中发挥重要作用。然而，大多数植物lncRNA的生物学功能仍然未知。我们之前发现了一个与番茄成熟相关的lncRNA，lncRNA 1459。在这里，我们克隆了全长lncRNA 1459，得到两个转录异构体。此外，lncRNA 1459在细胞核中表现出特定的位置。此外，为了充分鉴定lncRNA 1459在番茄成熟中的功能，使用成簇规则间隔短回文重复序列（CRISPR）/相关蛋白9（Cas9）诱导的基因组编辑技术开发了lncRNA 1459的功能丧失突变体。与野生型果实相比，lncRNA 1459突变体显著抑制了番茄的成熟过程。lncRNA 1459突变体的乙烯产生和番茄红素积累在很大程度上受到抑制。此外，与乙烯和类胡萝卜素生物合成相关的基因明显下调lncRNA 1459突变体相比，野生型水果。此外，当lncRNA 1459被敲除时，许多成熟相关基因的表达发生了显著变化。潜在的番茄成熟相关lncRNA的表达也发生了特异性变化后，敲除lncRNA 1459。综上所述，这些结果提供了深入了解lncRNA 1459在番茄果实成熟中的作用。
With the development of high-throughput sequencing, many long non-coding RNAs (lncRNAs) have been found to play important roles in diverse biological processes. However, the biological functions of most plant lncRNAs are still unknown. We have previously discovered a tomato ripening-related lncRNA, lncRNA1459. Here, we cloned the full-length lncRNA1459, giving two transcript isoforms. In addition, lncRNA1459 exhibited a specific location in the nucleus. Furthermore, in order to fully identify the function of lncRNA1459 in tomato ripening, loss-of-function mutants of lncRNA1459 were developed using clustered regularly interspaced short palindromic repeats (CRISPR)/-associated protein 9 (Cas9)-induced genome editing technology. Compared with wild-type fruits, the tomato ripening process was significantly repressed in lncRNA1459 mutants. Ethylene production and lycopene accumulation were largely repressed in lncRNA1459 mutants. Additionally, genes related to ethylene and carotenoid biosynthesis were distinctly downregulated in lncRNA1459 mutants compared with wild-type fruits. Moreover, expression of numerous ripening-related genes was changed significantly when lncRNA1459 was knocked out. Expression of potential tomato ripening-related lncRNAs was also specifically changed after knocking out lncRNA1459. Taken together, these results provide insight into the role of lncRNA1459 in tomato fruit ripening.