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Derepression of specific miRNA-target genes in rice using CRISPR/Cas9

MicroRNAs (miRNAs) target specific mRNA molecules based on sequence complementarity for their degradation or repression of translation, thereby regulating various developmental and physiological processes in eukaryotic organisms. Expressing the target mimicry (MIM) and short tandem target mimicry (S...

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Autores principales: Lin, Yarong, Zhu, Yiwang, Cui, Yuchao, Chen, Rui, Chen, Zaijie, Li, Gang, Fan, Meiying, Chen, Jianmin, Li, Yan, Guo, Xinrui, Zheng, Xijun, Chen, Liang, Wang, Feng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8547147/
https://www.ncbi.nlm.nih.gov/pubmed/34283216
http://dx.doi.org/10.1093/jxb/erab336
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author Lin, Yarong
Zhu, Yiwang
Cui, Yuchao
Chen, Rui
Chen, Zaijie
Li, Gang
Fan, Meiying
Chen, Jianmin
Li, Yan
Guo, Xinrui
Zheng, Xijun
Chen, Liang
Wang, Feng
author_facet Lin, Yarong
Zhu, Yiwang
Cui, Yuchao
Chen, Rui
Chen, Zaijie
Li, Gang
Fan, Meiying
Chen, Jianmin
Li, Yan
Guo, Xinrui
Zheng, Xijun
Chen, Liang
Wang, Feng
author_sort Lin, Yarong
collection PubMed
description MicroRNAs (miRNAs) target specific mRNA molecules based on sequence complementarity for their degradation or repression of translation, thereby regulating various developmental and physiological processes in eukaryotic organisms. Expressing the target mimicry (MIM) and short tandem target mimicry (STTM) can block endogenous activity of mature miRNAs and eliminate the inhibition of their target genes, resulting in phenotypic changes due to higher expression of the target genes. Here, we report a strategy to achieve derepression of interested miRNA-target genes through CRISPR/Cas9-based generation of in-frame mutants within the miRNA-complementary sequence of the target gene. We show that two rice genes, OsGRF4 (GROWTH REGULATING FACTOR 4) and OsGRF8 carrying in-frame mutants with disruption of the miR396 recognition sites, escape from miR396-mediated post-transcriptional silencing, resulting in enlarged grain size and increase in brown planthopper (BPH) resistance, in their respective transgenic rice lines. These results demonstrate that CRISPR/Cas9-mediated disruption of miRNA target sites can be effectively employed to precisely derepress particular target genes of functional importance for trait improvement in plants.
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spelling pubmed-85471472021-10-27 Derepression of specific miRNA-target genes in rice using CRISPR/Cas9 Lin, Yarong Zhu, Yiwang Cui, Yuchao Chen, Rui Chen, Zaijie Li, Gang Fan, Meiying Chen, Jianmin Li, Yan Guo, Xinrui Zheng, Xijun Chen, Liang Wang, Feng J Exp Bot Research Papers MicroRNAs (miRNAs) target specific mRNA molecules based on sequence complementarity for their degradation or repression of translation, thereby regulating various developmental and physiological processes in eukaryotic organisms. Expressing the target mimicry (MIM) and short tandem target mimicry (STTM) can block endogenous activity of mature miRNAs and eliminate the inhibition of their target genes, resulting in phenotypic changes due to higher expression of the target genes. Here, we report a strategy to achieve derepression of interested miRNA-target genes through CRISPR/Cas9-based generation of in-frame mutants within the miRNA-complementary sequence of the target gene. We show that two rice genes, OsGRF4 (GROWTH REGULATING FACTOR 4) and OsGRF8 carrying in-frame mutants with disruption of the miR396 recognition sites, escape from miR396-mediated post-transcriptional silencing, resulting in enlarged grain size and increase in brown planthopper (BPH) resistance, in their respective transgenic rice lines. These results demonstrate that CRISPR/Cas9-mediated disruption of miRNA target sites can be effectively employed to precisely derepress particular target genes of functional importance for trait improvement in plants. Oxford University Press 2021-07-20 /pmc/articles/PMC8547147/ /pubmed/34283216 http://dx.doi.org/10.1093/jxb/erab336 Text en © The Author(s) 2021. Published by Oxford University Press on behalf of the Society for Experimental Biology. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Papers
Lin, Yarong
Zhu, Yiwang
Cui, Yuchao
Chen, Rui
Chen, Zaijie
Li, Gang
Fan, Meiying
Chen, Jianmin
Li, Yan
Guo, Xinrui
Zheng, Xijun
Chen, Liang
Wang, Feng
Derepression of specific miRNA-target genes in rice using CRISPR/Cas9
title Derepression of specific miRNA-target genes in rice using CRISPR/Cas9
title_full Derepression of specific miRNA-target genes in rice using CRISPR/Cas9
title_fullStr Derepression of specific miRNA-target genes in rice using CRISPR/Cas9
title_full_unstemmed Derepression of specific miRNA-target genes in rice using CRISPR/Cas9
title_short Derepression of specific miRNA-target genes in rice using CRISPR/Cas9
title_sort derepression of specific mirna-target genes in rice using crispr/cas9
topic Research Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8547147/
https://www.ncbi.nlm.nih.gov/pubmed/34283216
http://dx.doi.org/10.1093/jxb/erab336
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