Large DNA fragment deletion in lncRNA77580 regulates neighboring gene expression in soybean (Glycine max)
Fengjuan Niu A , Qiyan Jiang
A * , Xianjun Sun A , Zheng Hu A , Lixia Wang A and Hui Zhang A *
+ Author Affiliations
- Author Affiliations
A Institute of Crop Science, National Key Facility for Crop Gene Resources and Genetic Improvement, Chinese Academy of Agricultural Sciences, Beijing, P. R. China.
Functional Plant Biology 48(11) 1139-1147 https://doi.org/10.1071/FP20400
Submitted: 22 December 2020 Accepted: 2 August 2021 Published: 29 September 2021
© 2021 The Author(s) (or their employer(s)). Published by CSIRO Publishing
Abstract
Long non-coding RNAs (lncRNAs) affect gene expressions via a wide range of mechanisms and are considered important regulators of numerous essential biological processes, including abiotic stress responses. However, the biological functions of most lncRNAs are yet to be determined. Moreover, to date, no effective methods have been developed to study the function of plant lncRNAs. We previously discovered a salt stress-related lncRNA, lncRNA77580 in soybean (Glycine max L.). In this study, we cloned the full-length lncRNA77580 and found that it shows nuclear-specific localisation. Furthermore, we employed CRISPR/Cas9 technology to induce large DNA fragment deletions in lncRNA77580 in soybean using a dual-single guide RNA/Cas9 design. As a result, we obtained deletion mutant soybean roots with targeted genomic fragment deletion in lncRNA77580. Deletion and overexpression of lncRNA77580 were found to alter the expression of several neighboring protein-coding genes associated with the response to salt stress. The longer the deleted DNA fragment in lncRNA77580, the greater the influence on the expression of lncRNA77580 itself and neighboring genes. Collectively, the findings of this study revealed that large DNA fragment deletion in lncRNAs using the CRISPR/Cas9 system is a powerful method to obtain functional mutations of soybean lncRNAs that could benefit future research on lncRNA function in soybean.
Keywords: CRISPR/Cas9, gene editing, gene expression, hairy root, large DNA fragment deletion, long non-coding RNA, salt stress, soybean.
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