Journal
JOURNAL OF EXPERIMENTAL BOTANY
Volume 67, Issue 21, Pages 6101-6110Publisher
OXFORD UNIV PRESS
DOI: 10.1093/jxb/erw371
Keywords
Centromere; chromatin; fluorescent protein; genome editing; live cell imaging; rDNA; telomere; transcription activator-like effector
Categories
Funding
- Japan Science and Technology Agency
- Ministry of Education, Culture, Sports, Science and Technology
- Japan Society for the Promotion of Science [JP25120726, JP26291067, JP15H05962, JP15H05955]
- Grants-in-Aid for Scientific Research [26291067, 15H05962, 16K18472] Funding Source: KAKEN
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Transcription activator-like effectors fused to fluorescent proteins can visualize repetitive genomic sequences including centromere, telomere, and rDNA sequences for analysing chromatin dynamics in living plant cells.Live imaging of the dynamics of nuclear organization provides the opportunity to uncover the mechanisms responsible for four-dimensional genome architecture. Here, we describe the use of fluorescent protein (FP) fusions of transcription activator-like effectors (TALEs) to visualize endogenous genomic sequences in Arabidopsis thaliana. The ability to engineer sequence-specific TALEs permits the investigation of precise genomic sequences. We could detect TALE-FP signals associated with centromeric, telomeric, and rDNA repeats and the signal distribution was consistent with that observed by fluorescent in situ hybridization. TALE-FPs are advantageous because they permit the observation of intact tissues. We used our TALE-FP method to investigate the nuclei of several multicellular plant tissues including roots, hypocotyls, leaves, and flowers. Because TALE-FPs permit live-cell imaging, we successfully observed the temporal dynamics of centromeres and telomeres in plant organs. Fusing TALEs to multimeric FPs enhanced the signal intensity when observing telomeres. We found that the mobility of telomeres was different in sub-nuclear regions. Transgenic plants stably expressing TALE-FPs will provide new insights into chromatin organization and dynamics in multicellular organisms.
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