An Efficient and Universal Protoplast Isolation Protocol Suitable for Transient Gene Expression Analysis and Single-Cell RNA Sequencing
文献类型：期刊论文
| 作者 | Wang, Juanjuan1; Wang, Yang1; Lu, Tianfeng; Yang, Xia; Liu, Jing; Dong, Yang1 ; Wang, Yinzheng1
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| 刊名 | INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
 |
| 出版日期 | 2022 |
| 卷号 | 23期号:7 |
| 关键词 | protoplast isolation cell walls Chirita pumila protocol transient gene expression single-cell RNA sequencing |
| DOI | 10.3390/ijms23073419 |
| 文献子类 | Article |
| 英文摘要 | The recent advent of single-cell RNA sequencing (scRNA-seq) has enabled access to the developmental landscape of a complex organ by monitoring the differentiation trajectory of every specialized cell type at the single-cell level. A main challenge in this endeavor is dissociating plant cells from the rigid cell walls and some species are recalcitrant to such cellular isolation. Here, we describe the establishment of a simple and efficient protocol for protoplast preparation in Chirita pumila, which includes two consecutive digestion processes with different enzymatic buffers. Using this protocol, we generated viable cell suspensions suitable for an array of expression analyses, including scRNA-seq. The universal application of this protocol was further tested by successfully isolating high-quality protoplasts from multiple organs (petals, fruits, tuberous roots, and gynophores) from representative species on the key branches of the angiosperm lineage. This work provides a robust method in plant science, overcoming barriers to isolating protoplasts in diverse plant species and opens a new avenue to study cell type specification, tissue function, and organ diversification in plants. |
| 学科主题 | Biochemistry & Molecular Biology ; Chemistry, Multidisciplinary |
| 出版地 | BASEL |
| 电子版国际标准刊号 | 1422-0067 |
| WOS关键词 | MESOPHYLL PROTOPLASTS ; ARABIDOPSIS ; PECTIN ; MORPHOGENESIS ; EVOLUTION ; SYSTEM |
| WOS研究方向 | Science Citation Index Expanded (SCI-EXPANDED) |
| 语种 | 英语 |
| WOS记录号 | WOS:000781113700001 |
| 出版者 | MDPI |
| 资助机构 | Natural Science Foundation of China [32170227, 31970239] |
| 源URL | [http://ir.ibcas.ac.cn/handle/2S10CLM1/28746]  |
| 专题 | 植物研究所_系统与进化植物学研究中心_系统与进化植物学研究中心_学位论文 |
| 作者单位 | 1.Chinese Acad Sci, Inst Bot, State Key Lab Systemat & Evolutionary Bot, Beijing 100093, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China |
| 推荐引用方式 GB/T 7714 | Wang, Juanjuan,Wang, Yang,Lu, Tianfeng,et al. An Efficient and Universal Protoplast Isolation Protocol Suitable for Transient Gene Expression Analysis and Single-Cell RNA Sequencing[J]. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES,2022,23(7). |
| APA | Wang, Juanjuan.,Wang, Yang.,Lu, Tianfeng.,Yang, Xia.,Liu, Jing.,...&Wang, Yinzheng.(2022).An Efficient and Universal Protoplast Isolation Protocol Suitable for Transient Gene Expression Analysis and Single-Cell RNA Sequencing.INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES,23(7). |
| MLA | Wang, Juanjuan,et al."An Efficient and Universal Protoplast Isolation Protocol Suitable for Transient Gene Expression Analysis and Single-Cell RNA Sequencing".INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES 23.7(2022). |
入库方式： OAI收割
来源：植物研究所
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