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 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.
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, Noda, Chiba 278-8510, Japan.
Satoru
S
Fujimoto
Graduate School of Science, Kyoto University, Kyoto 606-8502, Japan.|PRESTO, JST, Saitama 332-0012, Japan.
Shigeo S
SS
Sugano
Institute of Transformative Bio-Molecules, Nagoya University, Nagoya 464-8601, Japan.
Keiko
K
Kuwata
Faculty of Bioscience and Bioindustry, Tokushima University, Tokushima 770-8513, Japan.
Keishi
K
Osakabe
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, Noda, Chiba 278-8510, Japan sachi@rs.tus.ac.jp.
Sachihiro
S
Matsunaga
D017360:Arabidopsis / Q000235:genetics*; D018745:Genome, Plant / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D047468:Immunoprecipitation; D017404:In Situ Hybridization, Fluorescence; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics*; D000071336:Transcription Activator-Like Effectors / Q000235:genetics*
D000071336:Transcription Activator-Like Effectors; D049452:Green Fluorescent Proteins
Centromere; chromatin; fluorescent protein; genome editing; live cell imaging; rDNA; telomere; transcription activator-like effector.