2021
DOI: 10.1038/s41477-021-00976-0
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Quantitative imaging of RNA polymerase II activity in plants reveals the single-cell basis of tissue-wide transcriptional dynamics

Abstract: The responses of plants to their environment often hinge on the spatiotemporal dynamics of transcriptional regulation. While live-imaging tools have been used extensively to quantitatively capture rapid transcriptional dynamics in living animal cells, lack of implementation of these technologies in plants has limited concomitant quantitative studies. Here, we applied the PP7 and MS2 RNA-labeling technologies for the quantitative imaging of RNA polymerase II activity dynamics in single cells of living plants as… Show more

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Cited by 43 publications
(35 citation statements)
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“…Thus, these 'pathways' may not exist in any one cell suggesting heterogeneity in cell transcriptomes. Quantitative imaging of single cell transcriptional dynamics in plant cells have demonstrated that there are large differences between neighbouring cells, that is consistent with the conclusion that we have reached here with the comparison cell specific transcriptomes (Alamos et al, 2021;Hani et al, 2021).…”
Section: Discussionsupporting
confidence: 91%
“…Thus, these 'pathways' may not exist in any one cell suggesting heterogeneity in cell transcriptomes. Quantitative imaging of single cell transcriptional dynamics in plant cells have demonstrated that there are large differences between neighbouring cells, that is consistent with the conclusion that we have reached here with the comparison cell specific transcriptomes (Alamos et al, 2021;Hani et al, 2021).…”
Section: Discussionsupporting
confidence: 91%
“…Thus, the observed response is likely an average of many different scenarios that can blur interpretations. As single-cell approaches are picking up in plants, especially scRNA-seq [ 80 ], transcriptional cell-to-cell variability during development and in response exogenous stimulation is becoming more and more apparent [ 81 , 82 ]. Therefore, it is expected that the action of dCas9–CR fusions will also give heterogeneous outcomes that will have to be carefully monitored.…”
Section: Exploiting Current Limitations In Using Crispr/dcas9 Approaches and Going Beyondmentioning
confidence: 99%
“…We then generated reporter lines in which we could measure mRNA transcription in real time (Figure S12). The RPT2 promoter was used to drive transcription of a mRNA tagged in its 5' with a PP7 sequence recognized by a co-expressed GFP-tagged PP7 phage coat protein, enabling identification of nascent mRNA as fluorescent puncta (pRPT2:PP7 -Figure S12, (Larson et al, 2011;Alamos et al, 2021)). Imaging of dark-grown pRPT2:PP7 lines exposed to light was able to recapitulate light-induced transcriptional initiation (Figure 3A).…”
Section: Real-time Transcription Imaging Reveals An Earlier Timing Of Light-induced Transcription Initiationmentioning
confidence: 99%
“…To generate pRPT2:LUC lines, 3342 bp upstream of the RPT2 start codon were amplified by PCR with primers BamHI-pRPT2-5/ pRPT2-3-PstI and cloned into the pC1302-35S:RLUC vector (Zhang et al, 2013). To generate pRPT2:PP7 lines, 3356 bp upstream of the RPT2 start codon were amplified by PCR with primers 13Rb-RPT2F and 13Rb-RPT2R and cloned into the AL13Rb plasmid (Addgene # 161006) (Alamos et al, 2021). Primers are described in Supplementary Dataset S7.…”
Section: Plant Materials and Growth Conditionsmentioning
confidence: 99%
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