{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T18:45:19Z","timestamp":1792003519775,"version":"4.4.0"},"update-to":[{"DOI":"10.1371\/journal.pbio.3004043","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T00:00:00Z","timestamp":1791936000000}}],"reference-count":46,"publisher":"Public Library of Science (PLoS)","issue":"10","license":[{"start":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T00:00:00Z","timestamp":1791244800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32201241"],"award-info":[{"award-number":["32201241"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32171457"],"award-info":[{"award-number":["32171457"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100018542","name":"Natural Science Foundation of Sichuan Province","doi-asserted-by":"publisher","award":["25NSFSC2260"],"award-info":[{"award-number":["25NSFSC2260"]}],"id":[{"id":"10.13039\/501100018542","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100019448","name":"Zigong Science and Technology Program of China","doi-asserted-by":"publisher","award":["2024CDZG-21"],"award-info":[{"award-number":["2024CDZG-21"]}],"id":[{"id":"10.13039\/501100019448","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosbiology.org"],"crossmark-restriction":false},"short-container-title":["PLoS Biol"],"abstract":"<jats:p>\n                    Visualizing the spatiotemporal dynamics of native RNA molecules with single-transcript-level sensitivity in living plant cells remains a formidable challenge due to the high background noise caused by the unique cellular architecture of plants. Here, using transiently transformed\n                    <jats:italic>Nicotiana benthamiana<\/jats:italic>\n                    cells and stable\n                    <jats:italic>Arabidopsis thaliana<\/jats:italic>\n                    lines, we present a high-signal-to-noise RNA imaging platform that couples CRISPR\u2013dCas13X with a binding-associated fluorescence stabilization strategy. We identified a novel plant-specific degron that selectively targets unbound probes for proteasomal degradation while preserving target-associated signal in a manner consistent with binding-dependent protection, enabling high-fidelity detection of endogenous mRNA puncta with minimal perturbation. Leveraging this platform, we characterized dynamics of\n                    <jats:italic>FT<\/jats:italic>\n                    mRNA trafficking in living plant cells, including plasmodesmata-targeting, cell-to-cell trafficking, and long-distance movement. Loss-of-function analysis further identified the established RNA-binding protein GRP7 as a factor that contributes to efficient\n                    <jats:italic>FT<\/jats:italic>\n                    mRNA transport. This work establishes a useful toolkit for investigating plant RNA biology and provides mechanistic insight into the systemic transport of florigenic signals.\n                  <\/jats:p>","DOI":"10.1371\/journal.pbio.3004043","type":"journal-article","created":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T17:43:02Z","timestamp":1791308582000},"page":"e3004043","update-policy":"https:\/\/doi.org\/10.1371\/journal.pbio.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["A binding-activated CRISPR\/dCas13X imaging system enables precise RNA tracking in plants"],"prefix":"10.1371","volume":"24","author":[{"given":"Jiayu","family":"Zhang","sequence":"first","affiliation":[{"name":"Sichuan University","place":["Chengdu, China"],"department":["Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yiran","family":"Tao","sequence":"additional","affiliation":[{"name":"Sichuan University","place":["Chengdu, China"],"department":["Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yun","family":"Zhao","sequence":"additional","affiliation":[{"name":"Sichuan University","place":["Chengdu, China"],"department":["Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miaomiao","family":"Lei","sequence":"additional","affiliation":[{"name":"Sichuan University","place":["Chengdu, China"],"department":["Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yucheng","family":"Shen","sequence":"additional","affiliation":[{"name":"Sichuan University","place":["Chengdu, China"],"department":["Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qicong","family":"Li","sequence":"additional","affiliation":[{"name":"Sichuan University","place":["Chengdu, China"],"department":["Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chenxu","family":"Liu","sequence":"additional","affiliation":[{"name":"Sichuan University","place":["Chengdu, China"],"department":["Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3767-8291","authenticated-orcid":true,"given":"Jiuyuan","family":"Bai","sequence":"additional","affiliation":[{"name":"Sichuan University","place":["Chengdu, China"],"department":["Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,10,6]]},"reference":[{"issue":"8","key":"pbio.3004043.ref001","doi-asserted-by":"crossref","first-page":"1050","DOI":"10.1038\/s41477-021-00981-3","article-title":"Live single-cell transcriptional dynamics via RNA labelling during the phosphate response in plants","volume":"7","author":"S Hani","year":"2021","journal-title":"Nat Plants"},{"issue":"10","key":"pbio.3004043.ref002","doi-asserted-by":"crossref","first-page":"5071","DOI":"10.3390\/ijms22105071","article-title":"Single-molecule imaging in living plant cells: A methodological review","volume":"22","author":"A-Y Guo","year":"2021","journal-title":"Int J Mol Sci"},{"issue":"8","key":"pbio.3004043.ref003","doi-asserted-by":"crossref","first-page":"1037","DOI":"10.1038\/s41477-021-00976-0","article-title":"Quantitative imaging of RNA polymerase II activity in plants reveals the single-cell basis of tissue-wide transcriptional dynamics","volume":"7","author":"S Alamos","year":"2021","journal-title":"Nat Plants"},{"key":"pbio.3004043.ref004","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1080\/21541264.2023.2199669","article-title":"Understanding spatiotemporal coupling of gene expression using single molecule RNA imaging technologies","volume":"14","author":"A Gerber","year":"2023","journal-title":"Transcription"},{"key":"pbio.3004043.ref005","article-title":"Enhanced single RNA imaging reveals dynamic gene expression in live animals","volume":"12","author":"Y Hu","year":"2023","journal-title":"Elife"},{"issue":"2","key":"pbio.3004043.ref006","doi-asserted-by":"crossref","first-page":"358","DOI":"10.1016\/j.cell.2016.05.025","article-title":"Enhancer control of transcriptional bursting","volume":"166","author":"T Fukaya","year":"2016","journal-title":"Cell"},{"issue":"2","key":"pbio.3004043.ref007","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1038\/nrm3918","article-title":"In the right place at the right time: Visualizing and understanding mRNA localization","volume":"16","author":"AR Buxbaum","year":"2015","journal-title":"Nat Rev Mol Cell Biol"},{"issue":"14","key":"pbio.3004043.ref008","doi-asserted-by":"crossref","first-page":"3105","DOI":"10.1002\/cne.23073","article-title":"Arc mRNA docks precisely at the base of individual dendritic spines indicating the existence of a specialized microdomain for synapse-specific mRNA translation","volume":"520","author":"JL Dynes","year":"2012","journal-title":"J Comp Neurol"},{"issue":"1","key":"pbio.3004043.ref009","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1038\/s41592-025-02782-4","article-title":"RNA-stabilized coat proteins for sensitive and simultaneous imaging of distinct single mRNAs in live cells","volume":"23","author":"CJ Kuffner","year":"2026","journal-title":"Nat Methods"},{"issue":"4","key":"pbio.3004043.ref010","doi-asserted-by":"crossref","first-page":"437","DOI":"10.1016\/S1097-2765(00)80143-4","article-title":"Localization of ASH1 mRNA particles in living yeast","volume":"2","author":"E Bertrand","year":"1998","journal-title":"Mol Cell"},{"key":"pbio.3004043.ref011","doi-asserted-by":"crossref","first-page":"1216232","DOI":"10.3389\/fcell.2023.1216232","article-title":"Live cell imaging of DNA and RNA with fluorescent signal amplification and background reduction techniques","volume":"11","author":"S Lu","year":"2023","journal-title":"Front Cell Dev Biol"},{"issue":"3","key":"pbio.3004043.ref012","doi-asserted-by":"crossref","DOI":"10.1002\/wrna.1694","article-title":"Applications of the versatile CRISPR-Cas13 RNA targeting system","volume":"13","author":"M Kordy\u015b","year":"2022","journal-title":"Wiley Interdiscip Rev RNA"},{"issue":"13","key":"pbio.3004043.ref013","doi-asserted-by":"crossref","first-page":"2004685","DOI":"10.1002\/advs.202004685","article-title":"Functional features and current applications of the RNA-targeting type VI CRISPR-Cas systems","volume":"8","author":"V Per\u010dulija","year":"2021","journal-title":"Adv Sci (Weinh)"},{"issue":"17","key":"pbio.3004043.ref014","doi-asserted-by":"crossref","first-page":"2665","DOI":"10.3390\/cells11172665","article-title":"Applications of CRISPR\/Cas13-based RNA editing in plants","volume":"11","author":"NR Kavuri","year":"2022","journal-title":"Cells"},{"issue":"3","key":"pbio.3004043.ref015","doi-asserted-by":"crossref","first-page":"635","DOI":"10.1016\/j.cell.2014.09.039","article-title":"A protein-tagging system for signal amplification in gene expression and fluorescence imaging","volume":"159","author":"ME Tanenbaum","year":"2014","journal-title":"Cell"},{"issue":"4","key":"pbio.3004043.ref016","doi-asserted-by":"crossref","first-page":"1371","DOI":"10.1038\/s41596-019-0284-x","article-title":"Quantification of mRNA translation in live cells using single-molecule imaging","volume":"15","author":"D Khuperkar","year":"2020","journal-title":"Nat Protoc"},{"key":"pbio.3004043.ref017","doi-asserted-by":"crossref","first-page":"114074","DOI":"10.1016\/j.bios.2022.114074","article-title":"Live imaging of RNA and RNA splicing in mammalian cells via the dcas13a-SunTag-BiFC system","volume":"204","author":"M Chen","year":"2022","journal-title":"Biosens Bioelectron"},{"issue":"6","key":"pbio.3004043.ref018","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pgen.1010731","article-title":"Development of AlissAID system targeting GFP or mCherry fusion protein","volume":"19","author":"Y Ogawa","year":"2023","journal-title":"PLoS Genet"},{"issue":"5","key":"pbio.3004043.ref019","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1038\/s41589-018-0021-8","article-title":"The dTAG system for immediate and target-specific protein degradation","volume":"14","author":"B Nabet","year":"2018","journal-title":"Nat Chem Biol"},{"issue":"12","key":"pbio.3004043.ref020","doi-asserted-by":"crossref","first-page":"917","DOI":"10.1038\/nmeth.1401","article-title":"An auxin-based degron system for the rapid depletion of proteins in nonplant cells","volume":"6","author":"K Nishimura","year":"2009","journal-title":"Nat Methods"},{"key":"pbio.3004043.ref021","doi-asserted-by":"crossref","first-page":"866820","DOI":"10.3389\/fcell.2022.866820","article-title":"Progress of CRISPR-Cas13 mediated live-cell RNA imaging and detection of RNA-protein interactions","volume":"10","author":"H Cao","year":"2022","journal-title":"Front Cell Dev Biol"},{"issue":"47","key":"pbio.3004043.ref022","doi-asserted-by":"crossref","first-page":"14032","DOI":"10.1039\/D2SC04656C","article-title":"Live-cell RNA imaging using the CRISPR-dCas13 system with modified sgRNAs appended with fluorescent RNA aptamers","volume":"13","author":"H Tang","year":"2022","journal-title":"Chem Sci"},{"issue":"6","key":"pbio.3004043.ref023","doi-asserted-by":"crossref","DOI":"10.1016\/j.devcel.2019.02.022","article-title":"Spatiotemporal developmental trajectories in the Arabidopsis root revealed using high-throughput single-cell RNA sequencing","volume":"48","author":"T Denyer","year":"2019","journal-title":"Dev Cell"},{"issue":"4","key":"pbio.3004043.ref024","doi-asserted-by":"crossref","DOI":"10.1016\/j.devcel.2022.01.008","article-title":"A single-cell Arabidopsis root atlas reveals developmental trajectories in wild-type and cell identity mutants","volume":"57","author":"R Shahan","year":"2022","journal-title":"Dev Cell"},{"issue":"7","key":"pbio.3004043.ref025","doi-asserted-by":"crossref","DOI":"10.1016\/j.celrep.2019.04.054","article-title":"High-throughput single-cell transcriptome profiling of plant cell types","volume":"27","author":"CN Shulse","year":"2019","journal-title":"Cell Rep"},{"issue":"1","key":"pbio.3004043.ref026","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1186\/s13007-022-00849-3","article-title":"Development of a split fluorescent protein-based RNA live-cell imaging system to visualize mRNA distribution in plants","volume":"18","author":"N-C Huang","year":"2022","journal-title":"Plant Methods"},{"issue":"2","key":"pbio.3004043.ref027","doi-asserted-by":"crossref","first-page":"604","DOI":"10.1104\/pp.18.00107","article-title":"Selective targeting of mobile mRNAs to plasmodesmata for cell-to-cell movement","volume":"177","author":"K-R Luo","year":"2018","journal-title":"Plant Physiol"},{"issue":"6459","key":"pbio.3004043.ref028","doi-asserted-by":"crossref","first-page":"1301","DOI":"10.1126\/science.aax7852","article-title":"CRISPR-mediated live imaging of genome editing and transcription","volume":"365","author":"H Wang","year":"2019","journal-title":"Science"},{"issue":"6","key":"pbio.3004043.ref029","article-title":"Dynamic imaging of RNA in living cells by CRISPR-Cas13 systems","volume":"76","author":"L-Z Yang","year":"2019","journal-title":"Mol Cell"},{"issue":"5","key":"pbio.3004043.ref030","doi-asserted-by":"crossref","first-page":"499","DOI":"10.1038\/s41592-021-01124-4","article-title":"Programmable RNA editing with compact CRISPR-Cas13 systems from uncultivated microbes","volume":"18","author":"C Xu","year":"2021","journal-title":"Nat Methods"},{"issue":"9","key":"pbio.3004043.ref031","doi-asserted-by":"crossref","first-page":"1256","DOI":"10.1038\/s41587-022-01649-9","article-title":"Precise transcript targeting by CRISPR-Csm complexes","volume":"41","author":"D Colognori","year":"2023","journal-title":"Nat Biotechnol"},{"issue":"12","key":"pbio.3004043.ref032","doi-asserted-by":"crossref","first-page":"2023","DOI":"10.1038\/s41587-024-02540-5","article-title":"Single-molecule live-cell RNA imaging with CRISPR-Csm","volume":"43","author":"C Xia","year":"2025","journal-title":"Nat Biotechnol"},{"issue":"8","key":"pbio.3004043.ref033","doi-asserted-by":"crossref","DOI":"10.1002\/cpz1.219","article-title":"Targeted Protein depletion using the auxin-inducible degron 2 (AID2) system","volume":"1","author":"Y Saito","year":"2021","journal-title":"Curr Protoc"},{"issue":"1","key":"pbio.3004043.ref034","doi-asserted-by":"crossref","first-page":"10738","DOI":"10.1038\/s41467-025-65768-y","article-title":"Programmable targeted RNA degradation via dCas13d-directed chaperone-mediated autophagy (dCasCMA)","volume":"16","author":"H-P Wen","year":"2025","journal-title":"Nat Commun"},{"issue":"39","key":"pbio.3004043.ref035","article-title":"CRISPR\/dCas13(Rx) derived RNA N6-methyladenosine (m6A) dynamic modification in plant","volume":"11","author":"L Yu","year":"2024","journal-title":"Adv Sci (Weinh)"},{"issue":"5827","key":"pbio.3004043.ref036","doi-asserted-by":"crossref","first-page":"1030","DOI":"10.1126\/science.1141752","article-title":"FT protein movement contributes to long-distance signaling in floral induction of Arabidopsis","volume":"316","author":"L Corbesier","year":"2007","journal-title":"Science"},{"issue":"5827","key":"pbio.3004043.ref037","doi-asserted-by":"crossref","first-page":"1033","DOI":"10.1126\/science.1141753","article-title":"Hd3a protein is a mobile flowering signal in rice","volume":"316","author":"S Tamaki","year":"2007","journal-title":"Science"},{"issue":"5","key":"pbio.3004043.ref038","doi-asserted-by":"crossref","first-page":"653","DOI":"10.4161\/rna.19965","article-title":"Long-distance movement of Arabidopsis FLOWERING LOCUS T RNA participates in systemic floral regulation","volume":"9","author":"K-J Lu","year":"2012","journal-title":"RNA Biol"},{"issue":"1","key":"pbio.3004043.ref039","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1186\/s13059-017-1332-x","article-title":"Adaptation of iCLIP to plants determines the binding landscape of the clock-regulated RNA-binding protein AtGRP7","volume":"18","author":"K Meyer","year":"2017","journal-title":"Genome Biol"},{"issue":"9","key":"pbio.3004043.ref040","doi-asserted-by":"crossref","first-page":"2040","DOI":"10.1093\/pcp\/pcz124","article-title":"Regulation of flowering time by the RNA-binding proteins AtGRP7 and AtGRP8","volume":"60","author":"A Steffen","year":"2019","journal-title":"Plant Cell Physiol"},{"issue":"1","key":"pbio.3004043.ref041","doi-asserted-by":"crossref","first-page":"16018","DOI":"10.1038\/s41598-024-66955-5","article-title":"The glycine-rich domain of GRP7 plays a crucial role in binding long RNAs and facilitating phase separation","volume":"14","author":"KL L\u00fchmann","year":"2024","journal-title":"Sci Rep"},{"issue":"2","key":"pbio.3004043.ref042","doi-asserted-by":"crossref","first-page":"321","DOI":"10.1016\/j.molp.2019.12.001","article-title":"A plant SMALL RNA-BINDING PROTEIN 1 family mediates cell-to-cell trafficking of RNAi signals","volume":"13","author":"Y Yan","year":"2020","journal-title":"Mol Plant"},{"issue":"4","key":"pbio.3004043.ref043","doi-asserted-by":"crossref","first-page":"683","DOI":"10.1111\/tpj.12662","article-title":"Systemic delivery of siRNA in pumpkin by a plant PHLOEM SMALL RNA-BINDING PROTEIN 1-ribonucleoprotein complex","volume":"80","author":"BK Ham","year":"2014","journal-title":"Plant J"},{"issue":"2","key":"pbio.3004043.ref044","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1111\/tpj.16326","article-title":"Arabidopsis callus grafts: A system to study symplasmic intercellular macromolecular transport and gene silencing spread","volume":"115","author":"Y Hasbio\u011flu","year":"2023","journal-title":"Plant J"},{"issue":"7","key":"pbio.3004043.ref045","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1038\/nmeth.2019","article-title":"Fiji: An open-source platform for biological-image analysis","volume":"9","author":"J Schindelin","year":"2012","journal-title":"Nat Methods"},{"key":"pbio.3004043.ref046","doi-asserted-by":"crossref","first-page":"80","DOI":"10.1016\/j.ymeth.2016.09.016","article-title":"TrackMate: An open and extensible platform for single-particle tracking","volume":"115","author":"J-Y Tinevez","year":"2017","journal-title":"Methods"}],"updated-by":[{"DOI":"10.1371\/journal.pbio.3004043","type":"new_version","label":"New version","source":"publisher","updated":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T00:00:00Z","timestamp":1791936000000}}],"container-title":["PLOS Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pbio.3004043","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T17:52:09Z","timestamp":1792000329000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pbio.3004043"}},"subtitle":[],"editor":[{"given":"Simon","family":"Bullock","sequence":"first","affiliation":[{"name":"MRC Laboratory of Molecular Biology","place":["UNITED KINGDOM OF GREAT BRITAIN AND NORTHERN IRELAND"]}],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,10,6]]},"references-count":46,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2026,10,6]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pbio.3004043","relation":{},"ISSN":["1545-7885"],"issn-type":[{"value":"1545-7885","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,10,6]]}}}