水稻OsSm rna结合基因的CRISPR/Cas9突变资源

IF 5.7 1区 生物学 Q1 PLANT SCIENCES
Xiongxia Jin, Chao Ouyang, Tong Sun, Cong Li, Jinbao Gu, Baoguang An, Zhenyu Wang
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引用次数: 0

摘要

Pre-mRNA由真核生物DNA转录产生,通过剪接体剪切,去除内含子并连接外显子形成mRNA。剪接体是一个巨大的、高度动态的分子机器。其核心成分包括五种小核核糖核蛋白(snRNPs)和各种剪接体相关蛋白。保守的Smith (Sm)复合物和Sm样蛋白(LSm)是snRNPs的主要成分。Sm蛋白参与pre-mRNA剪接和mRNA降解等过程,调控基因表达,从而影响植物的生长发育和胁迫反应。虽然在水稻中发现了25种Sm蛋白,但它们在调节水稻生长发育中的具体作用尚不清楚。在本研究中,我们利用CRISPR/Cas9系统编辑了15个OsSm基因,获得了13个突变体,突变率在20.83% ~ 83.87%之间。与野生型(WT)相比,突变体表现出矮化,分蘖数减少,结实率或不育性降低,对疾病的易感性增加。一个Sm突变体ossmf-2表现出矮化、延迟开花和小粒。通过转录组分析,鉴定出OsMRG702、OsRGG2和OsLA1三个靶基因。OsSmF蛋白的突变可能导致这些基因的异常剪接,最终导致生长发育的抑制。我们的研究首先编辑了OsSm基因,并在水稻中建立了一个突变文库。大多数突变体表现出异常的生长发育,强调了OsSm蛋白在水稻生理中的重要作用。此外,这项工作解决了水稻中Sm蛋白功能表征的关键空白。由此产生的突变体集合提供了宝贵的种质资源,并为阐明剪接体组分及其靶基因在控制作物生长、发育和繁殖中的分子调控网络奠定了理论基础。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

A CRISPR/Cas9 mutant resource for OsSm RNA-binding genes in rice

A CRISPR/Cas9 mutant resource for OsSm RNA-binding genes in rice

Pre-mRNA, produced by eukaryotic DNA transcription, undergoes splicing by the spliceosome, which removes introns and joins exons to form mRNA. The spliceosome is a large and highly dynamic molecular machine. Its core components include five small nuclear ribonucleoproteins (snRNPs) and the various spliceosome-related proteins. The conserved Smith (Sm) complex and the Sm-like proteins (LSm) serve as primary components of the snRNPs. Sm proteins are involved in processes such as pre-mRNA splicing and mRNA degradation, which can regulate gene expression, thereby influencing plant growth, development, and stress responses. While 25 Sm proteins have been identified in rice, their specific roles in regulating rice growth and development remain unclear. In this study, we employed the CRISPR/Cas9 system to edit 15 OsSm genes, and 13 mutants were obtained, with mutation rates ranging from 20.83 to 83.87%. In comparison to the wild type (WT), the mutants exhibited dwarfism, reduced tiller numbers, lower seed-setting rates or sterility, and increased susceptibility to diseases. One Sm mutant, ossmf-2, exhibited dwarfism, delayed flowering, and small grains. Through transcriptome analysis, three target genes, OsMRG702, OsRGG2, and OsLA1, were identified. Mutations of the OsSmF protein may lead to the abnormal splicing of these genes and finally lead to the inhibition of growth and development. Our study first edited the OsSm genes and generated a mutant library in rice. Most of the mutants exhibited abnormal growth and development, underscoring the essential roles of OsSm proteins in rice physiology. Furthermore, this work addresses a critical gap in the functional characterization of Sm proteins in rice. The resulting mutant collection offers valuable germplasm resources and lays a theoretical foundation for elucidating the molecular regulatory networks involving spliceosomal components and their target genes in the control of crop growth, development, and reproduction.

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来源期刊
The Plant Journal
The Plant Journal 生物-植物科学
CiteScore
13.10
自引率
4.20%
发文量
415
审稿时长
2.3 months
期刊介绍: Publishing the best original research papers in all key areas of modern plant biology from the world"s leading laboratories, The Plant Journal provides a dynamic forum for this ever growing international research community. Plant science research is now at the forefront of research in the biological sciences, with breakthroughs in our understanding of fundamental processes in plants matching those in other organisms. The impact of molecular genetics and the availability of model and crop species can be seen in all aspects of plant biology. For publication in The Plant Journal the research must provide a highly significant new contribution to our understanding of plants and be of general interest to the plant science community.
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