ArticlePlant methods2023
Establishment of an efficient cotton root protoplast isolation protocol suitable for single-cell RNA sequencing and transient gene expression analysis.
Article in Plant methods, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed, 18 citations in OpenAlex.
- An efficient method for tissue-specific protoplast isolation suitable for single-cell RNA sequencing and transient gene expression analysis in saffron (Crocus sativus L.).BMC biotechnology · 2026Article
- Genome Editing Approaches in Flax (International journal of molecular sciences · 2026Review
- Establishment of a protoplast isolation and transient transformation system for tung tree (Vernicia fordii).Plant methods · 2026Article
- Single-cell RNA-sequencing profiles reveal the developmental landscape of hawthorn leaves.BMC plant biology · 2026Article
- Protoplast isolation and transient gene expression in Suaeda aralocaspica, a halophyte with single-cell CPlant methods · 2026Article
- Identification of Phloem Cell Transcriptome via Single-Cell RNA-Sequencing.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Beyond a few bases: methods for large DNA insertion and gene targeting in plants.The Plant journal : for cell and molecular biology · 2025Review
- Isolation of Haustorium Protoplasts Optimized by Orthogonal Design for Transient Gene Expression inPlants (Basel, Switzerland) · 2024Article
- Expanding the targeting scope of CRISPR/Cas9-mediated genome editing by Cas9 variants inaBIOTECH · 2024Article
- Isolation, Purification, and Application of Protoplasts and Transient Expression Systems in Plants.International journal of molecular sciences · 2023Review
Corrections and comments
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Authors and funding
9 authors at 1 institution in 1 country.
Funding
Abstract
backgroundCotton has tremendous economic value worldwide; however, its allopolyploid nature and time-consuming transformation methods have hampered the development of cotton functional genomics. The protoplast system has proven to be an important and versatile tool for functional genomics, tissue-specific marker gene identification, tracking developmental trajectories, and genome editing in plants. Nevertheless, the isolation of abundant viable protoplasts suitable for single-cell RNA sequencing (scRNA-seq) and genome editing remains a challenge in cotton.
resultsWe established an efficient transient gene expression system using protoplasts isolated from cotton taproots. The system enables the isolation of large numbers of viable protoplasts and uses an optimized PEG-mediated transfection protocol. The highest yield (3.55 × 10
conclusionsThe protoplast isolation and transfection protocol developed in this study is stable, versatile, and time-saving. It will accelerate functional genomics and molecular breeding in cotton.
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