FlsnRNA-seq: protoplasting-free full-length single-nucleus RNA profiling in plants
Open Access
- 19 February 2021
- journal article
- research article
- Published by Springer Science and Business Media LLC in Genome Biology
- Vol. 22 (1), 1-14
- https://doi.org/10.1186/s13059-021-02288-0
Abstract
The broad application of single-cell RNA profiling in plants has been hindered by the prerequisite of protoplasting that requires digesting the cell walls from different types of plant tissues. Here, we present a protoplasting-free approach, flsnRNA-seq, for large-scale full-length RNA profiling at a single-nucleus level in plants using isolated nuclei. Combined with 10x Genomics and Nanopore long-read sequencing, we validate the robustness of this approach in Arabidopsis root cells and the developing endosperm. Sequencing results demonstrate that it allows for uncovering alternative splicing and polyadenylation-related RNA isoform information at the single-cell level, which facilitates characterizing cell identities.Keywords
Funding Information
- Key Laboratory of Molecular Design for Plant Cell Factory of Guangdong Higher Education Institutes (2019KSYS006)
- The Shenzhen Sci-Tech Fund (KYTDPT20181011104005)
- The Program for Guangdong Introducing Innovative and Entrepreneurial Teams (2016ZT06S172)
- the National key R&D Program of China Grant (2019YFA0903903)
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