Repositorium
Efficient Generation of diRNAs Requires Components in the Posttranscriptional Gene Silencing Pathway
462
Journal Article / 2017
Miki, Daisuke; Zhu, Peiying; Zhang, Wencan; Mao, Yanfei; Feng, Zhengyan; Huang, Huan; Zhang, Hui; Li, Yanqiang; Liu, Renyi; Zhang, Huiming; Qi, Yijun; Zhu, Jian-Kang
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.
Techniques
ID | Corresponding Author Country |
Plant Species | GE Technique Sequence Identifier |
Trait Type of Alteration |
Progress in Research Key Topic |
---|---|---|---|---|---|
1132 |
Miki, Daisuke China |
Arabidopsis thaliana |
CRISPR/Cas9 GUS |
No information SDN1 |
Basic research Basic research |
1133 |
Miki, Daisuke China |
Arabidopsis thaliana |
CRISPR/Cas9 GAI |
No information SDN1 |
Basic research Basic research |
1134 |
Miki, Daisuke China |
Arabidopsis thaliana |
CRISPR/Cas9 GL2 |
No information SDN1 |
Basic research Basic research |
1135 |
Miki, Daisuke China |
Arabidopsis thaliana |
CRISPR/Cas9 At5g36250 |
No information SDN1 |
Basic research Basic research |
1136 |
Miki, Daisuke China |
Arabidopsis thaliana |
CRISPR/Cas9 At2g36490 |
No information SDN1 |
Basic research Basic research |
1137 |
Miki, Daisuke China |
Arabidopsis thaliana |
CRISPR/Cas9 At5g04560 |
No information SDN1 |
Basic research Basic research |
1138 |
Miki, Daisuke China |
Arabidopsis thaliana |
CRISPR/Cas9 At1g31290 |
No information SDN1 |
Basic research Basic research |
1139 |
Miki, Daisuke China |
Arabidopsis thaliana |
CRISPR/Cas9 At5g54700 |
No information SDN1 |
Basic research Basic research |
1140 |
Miki, Daisuke China |
Oryza sativa |
CRISPR/Cas9 Pms3 |
No information SDN1 |
Basic research Basic research |
1141 |
Miki, Daisuke China |
Oryza sativa |
CRISPR/Cas9 PDS |
Albino phenotype SDN1 |
Basic research Basic research |
1142 |
Miki, Daisuke China |
Oryza sativa |
CRISPR/Cas9 MYB5 |
No information SDN1 |
Basic research Basic research |
1143 |
Miki, Daisuke China |
Oryza sativa |
CRISPR/Cas9 DERF1 |
No information SDN1 |
Basic research Basic research |
1144 |
Miki, Daisuke China |
Oryza sativa |
TALENs MST7 |
No information SDN1 |
Basic research Basic research |