A binding-activated CRISPR/dCas13X imaging system enables precise RNA tracking in plants
- 1Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University, Chengdu, China.
Abstract
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 Nicotiana benthamiana cells and stable Arabidopsis thaliana lines, we present a high-signal-to-noise RNA imaging platform that couples CRISPR–dCas13X 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 FT 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 FT mRNA transport. This work establishes a useful toolkit for investigating plant RNA biology and provides mechanistic insight into the systemic transport of florigenic signals.
Visualizing native RNA dynamics in living plant cells at single transcript resolution is limited by high background fluorescence. This study develops a high signal-to-noise CRISPR-dCas13X imaging platform that overcomes this barrier and provides insights into mRNA trafficking dynamics.
Related
- A novel CD69 binding ankyrin repeat protein (Ankyron®) enables detection of early activated and antigen-specific T cells in pigs
- Twin-prime editing enables endogenous protein visualization and mutant allele tracking
- AAVolve: Concatenated long-read deep sequencing enables whole capsid tracking during shuffled AAV library selection
- Dual‐Organelle Secretome Conjugation and Organ‐Uptake Tracking (DuO‐SCOUT) Enables Deep and Sensitive Mapping of the Secreted Proteins
- Dopamine Release from the Locus Coeruleus Enables Temporal Binding in the Hippocampus
BibTeX
@article{zhang2026bindingactivated,
title = {{A binding-activated CRISPR/dCas13X imaging system enables precise RNA tracking in plants}},
author = {Jiayu Zhang and Yiran Tao and Yun Zhao and Miaomiao Lei and Yucheng Shen and Qicong Li and Chenxu Liu and Jiuyuan Bai},
journal = {PLOS Biology},
year = {2026},
month = oct,
doi = {10.1371/journal.pbio.3004043},
url = {https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.3004043},
note = {CC BY},
}
RIS
TY - JOUR
TI - A binding-activated CRISPR/dCas13X imaging system enables precise RNA tracking in plants
AU - Jiayu Zhang
AU - Yiran Tao
AU - Yun Zhao
AU - Miaomiao Lei
AU - Yucheng Shen
AU - Qicong Li
AU - Chenxu Liu
AU - Jiuyuan Bai
PY - 2026
DA - 2026/10/06
JO - PLOS Biology
DO - 10.1371/journal.pbio.3004043
UR - https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.3004043
AB - 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 Nicotiana benthamiana cells and stable Arabidopsis thaliana lines, we present a high-signal-to-noise RNA imaging platform that couples CRISPR–dCas13X 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 FT 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 FT mRNA transport. This work establishes a useful toolkit for investigating plant RNA biology and provides mechanistic insight into the systemic transport of florigenic signals. Visualizing native RNA dynamics in living plant cells at single transcript resolution is limited by high background fluorescence. This study develops a high signal-to-noise CRISPR-dCas13X imaging platform that overcomes this barrier and provides insights into mRNA trafficking dynamics.
SN - 1545-7885
SN - 1544-9173
ER -