期刊
PLANT CELL
卷 33, 期 4, 页码 1135-1150出版社
OXFORD UNIV PRESS INC
DOI: 10.1093/plcell/koab020
关键词
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资金
- Science Foundation Ireland [13/IA/1820]
- Irish Research Council [GOIPG/2018/1960]
- Cost Action Impacts of Chromatin Domains on Plant Phenotypes (INDEPTH) [CA16212]
- EMBO Short Term Fellowship [8312]
- Science Foundation Ireland (SFI) [13/IA/1820] Funding Source: Science Foundation Ireland (SFI)
In this study, the researchers used Cas9-mediated genome editing in Arabidopsis to reduce the copy number of 45S rRNA genes to around 10%, creating low copy number (LCN) lines. Despite the significant reduction in rDNA copies, the transcriptional rates and steady-state levels of rRNA remained the same as wild-type plants. Furthermore, editing rRNA repeats using Cas9 technology provides insights into the mechanisms of rDNA dosage compensation and the impacts of low rDNA copy number on genome stability, development, and cellular processes.
The 45S rRNA genes (rDNA) are among the largest repetitive elements in eukaryotic genomes. rDNA consists of tandem arrays of rRNA genes, many of which are transcriptionally silenced. Silent rDNA repeats may act as 'back-up' copies for ribosome biogenesis and have nuclear organization roles. Through Cas9-mediated genome editing in the Arabidopsis thaliana female gametophyte, we reduced 45S rDNA copy number (CN) to a plateau of similar to 10%. Two independent lines had rDNA CNs reduced by up to 90% at the T7 generation, named low copy number (LCN) lines. Despite drastic reduction of rDNA copies, rRNA transcriptional rates, and steady-state levels remained the same as wild-type plants. Gene dosage compensation of rRNA transcript levels was associated with reduction of silencing histone marks at rDNA loci and altered Nucleolar Organiser Region 2 organization. Although overall genome integrity of LCN lines appears unaffected, a chromosome segmental duplication occurred in one of the lines. Transcriptome analysis of LCN seedlings identified several shared dysregulated genes and pathways in both independent lines. Cas9 genome editing of rRNA repeats to generate LCN lines provides a powerful technique to elucidate rDNA dosage compensation mechanisms and impacts of low rDNA CN on genome stability, development, and cellular processes.
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