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Visualization of specific repetitive genomic sequences with fluorescent TALEs in Arabidopsis thaliana

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 genomi...

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Autores principales: Fujimoto, Satoru, Sugano, Shigeo S., Kuwata, Keiko, Osakabe, Keishi, Matsunaga, Sachihiro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5100022/
https://www.ncbi.nlm.nih.gov/pubmed/27811079
http://dx.doi.org/10.1093/jxb/erw371
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author Fujimoto, Satoru
Sugano, Shigeo S.
Kuwata, Keiko
Osakabe, Keishi
Matsunaga, Sachihiro
author_facet Fujimoto, Satoru
Sugano, Shigeo S.
Kuwata, Keiko
Osakabe, Keishi
Matsunaga, Sachihiro
author_sort Fujimoto, Satoru
collection PubMed
description 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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spelling pubmed-51000222016-11-10 Visualization of specific repetitive genomic sequences with fluorescent TALEs in Arabidopsis thaliana Fujimoto, Satoru Sugano, Shigeo S. Kuwata, Keiko Osakabe, Keishi Matsunaga, Sachihiro J Exp Bot Research Paper 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. Oxford University Press 2016-11 2016-10-06 /pmc/articles/PMC5100022/ /pubmed/27811079 http://dx.doi.org/10.1093/jxb/erw371 Text en © The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology. http://creativecommons.org/licenses/by/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Paper
Fujimoto, Satoru
Sugano, Shigeo S.
Kuwata, Keiko
Osakabe, Keishi
Matsunaga, Sachihiro
Visualization of specific repetitive genomic sequences with fluorescent TALEs in Arabidopsis thaliana
title Visualization of specific repetitive genomic sequences with fluorescent TALEs in Arabidopsis thaliana
title_full Visualization of specific repetitive genomic sequences with fluorescent TALEs in Arabidopsis thaliana
title_fullStr Visualization of specific repetitive genomic sequences with fluorescent TALEs in Arabidopsis thaliana
title_full_unstemmed Visualization of specific repetitive genomic sequences with fluorescent TALEs in Arabidopsis thaliana
title_short Visualization of specific repetitive genomic sequences with fluorescent TALEs in Arabidopsis thaliana
title_sort visualization of specific repetitive genomic sequences with fluorescent tales in arabidopsis thaliana
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5100022/
https://www.ncbi.nlm.nih.gov/pubmed/27811079
http://dx.doi.org/10.1093/jxb/erw371
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