Efficient Generation of diRNAs Requires Components in the Posttranscriptional Gene Silencing Pathway
Abstract It has been reported that double-stranded break (DSB)-induced small RNAs (diRNAs) are generated via the RNA-directed DNA methylation pathway and function in DSB repair in Arabidposis. However, important questions remain regarding the biogenesis and function of diRNAs. Here, we used CRISPR/C...
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Nature Portfolio
2017
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oai:doaj.org-article:832b4b6f88db40e49e7e863c3ee5b1ac2021-12-02T15:04:57ZEfficient Generation of diRNAs Requires Components in the Posttranscriptional Gene Silencing Pathway10.1038/s41598-017-00374-72045-2322https://doaj.org/article/832b4b6f88db40e49e7e863c3ee5b1ac2017-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-00374-7https://doaj.org/toc/2045-2322Abstract It has been reported that double-stranded break (DSB)-induced small RNAs (diRNAs) are generated via the RNA-directed DNA methylation pathway and function in DSB repair in Arabidposis. However, important questions remain regarding the biogenesis and function of diRNAs. Here, we used CRISPR/Cas9- or TALEN-triggered DSBs to characterize diRNAs in Arabidopsis and rice. We found that 21-nt diRNAs were generated from a 35S promoter::GU-US reporter transgene targeted by CRISPR/Cas9. Unexpectedly, Pol II transcription of the transgene was required for efficient diRNA production and the level of diRNA accumulation correlated with the expression level of the transgene. diRNAs were not detected from CRISPR/Cas9- or TALEN-induced DSBs within the examined endogenous genes in Arabidopsis or rice. We also found that DCL4 and RDR6 that are known to be involved in posttranscriptional gene silencing were required to generate diRNAs. Our results suggest that DSBs are necessary but not sufficient for efficient diRNA generation and a high level of diRNAs is not necessary for DSB repair.Daisuke MikiPeiying ZhuWencan ZhangYanfei MaoZhengyan FengHuan HuangHui ZhangYanqiang LiRenyi LiuHuiming ZhangYijun QiJian-Kang ZhuNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017) |
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EN |
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Medicine R Science Q |
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Medicine R Science Q Daisuke Miki Peiying Zhu Wencan Zhang Yanfei Mao Zhengyan Feng Huan Huang Hui Zhang Yanqiang Li Renyi Liu Huiming Zhang Yijun Qi Jian-Kang Zhu Efficient Generation of diRNAs Requires Components in the Posttranscriptional Gene Silencing Pathway |
| description |
Abstract It has been reported that double-stranded break (DSB)-induced small RNAs (diRNAs) are generated via the RNA-directed DNA methylation pathway and function in DSB repair in Arabidposis. However, important questions remain regarding the biogenesis and function of diRNAs. Here, we used CRISPR/Cas9- or TALEN-triggered DSBs to characterize diRNAs in Arabidopsis and rice. We found that 21-nt diRNAs were generated from a 35S promoter::GU-US reporter transgene targeted by CRISPR/Cas9. Unexpectedly, Pol II transcription of the transgene was required for efficient diRNA production and the level of diRNA accumulation correlated with the expression level of the transgene. diRNAs were not detected from CRISPR/Cas9- or TALEN-induced DSBs within the examined endogenous genes in Arabidopsis or rice. We also found that DCL4 and RDR6 that are known to be involved in posttranscriptional gene silencing were required to generate diRNAs. Our results suggest that DSBs are necessary but not sufficient for efficient diRNA generation and a high level of diRNAs is not necessary for DSB repair. |
| format |
article |
| author |
Daisuke Miki Peiying Zhu Wencan Zhang Yanfei Mao Zhengyan Feng Huan Huang Hui Zhang Yanqiang Li Renyi Liu Huiming Zhang Yijun Qi Jian-Kang Zhu |
| author_facet |
Daisuke Miki Peiying Zhu Wencan Zhang Yanfei Mao Zhengyan Feng Huan Huang Hui Zhang Yanqiang Li Renyi Liu Huiming Zhang Yijun Qi Jian-Kang Zhu |
| author_sort |
Daisuke Miki |
| title |
Efficient Generation of diRNAs Requires Components in the Posttranscriptional Gene Silencing Pathway |
| title_short |
Efficient Generation of diRNAs Requires Components in the Posttranscriptional Gene Silencing Pathway |
| title_full |
Efficient Generation of diRNAs Requires Components in the Posttranscriptional Gene Silencing Pathway |
| title_fullStr |
Efficient Generation of diRNAs Requires Components in the Posttranscriptional Gene Silencing Pathway |
| title_full_unstemmed |
Efficient Generation of diRNAs Requires Components in the Posttranscriptional Gene Silencing Pathway |
| title_sort |
efficient generation of dirnas requires components in the posttranscriptional gene silencing pathway |
| publisher |
Nature Portfolio |
| publishDate |
2017 |
| url |
https://doaj.org/article/832b4b6f88db40e49e7e863c3ee5b1ac |
| work_keys_str_mv |
AT daisukemiki efficientgenerationofdirnasrequirescomponentsintheposttranscriptionalgenesilencingpathway AT peiyingzhu efficientgenerationofdirnasrequirescomponentsintheposttranscriptionalgenesilencingpathway AT wencanzhang efficientgenerationofdirnasrequirescomponentsintheposttranscriptionalgenesilencingpathway AT yanfeimao efficientgenerationofdirnasrequirescomponentsintheposttranscriptionalgenesilencingpathway AT zhengyanfeng efficientgenerationofdirnasrequirescomponentsintheposttranscriptionalgenesilencingpathway AT huanhuang efficientgenerationofdirnasrequirescomponentsintheposttranscriptionalgenesilencingpathway AT huizhang efficientgenerationofdirnasrequirescomponentsintheposttranscriptionalgenesilencingpathway AT yanqiangli efficientgenerationofdirnasrequirescomponentsintheposttranscriptionalgenesilencingpathway AT renyiliu efficientgenerationofdirnasrequirescomponentsintheposttranscriptionalgenesilencingpathway AT huimingzhang efficientgenerationofdirnasrequirescomponentsintheposttranscriptionalgenesilencingpathway AT yijunqi efficientgenerationofdirnasrequirescomponentsintheposttranscriptionalgenesilencingpathway AT jiankangzhu efficientgenerationofdirnasrequirescomponentsintheposttranscriptionalgenesilencingpathway |
| _version_ |
1718388966178160640 |