摘要
同一个基因在不同的细胞中表达存在一定的差异,常规的转录组测序方法(RNA-Seq)在组织或器官水平上进行分析,容易忽略细胞间的差异,但近几年迅速发展的高通量单细胞转录组测序技术在单细胞水平上分析,可以补充常规测序所忽略的异质性.目前,植物生物学领域已经全面开始单细胞转录组学,多组学研究成为趋势.本文综述了单细胞测序发展历程以及分离单细胞的方法并总结了各种方法的优缺点,介绍了近年来单细胞转录组测序技术在揭示植物细胞发育和分化过程、生物合成途径、应对生物与非生物胁迫的响应及在分析不同物种间细胞类型分化等方面取得的显著进展.此外,多组学联合应用不仅有助于人们更深入地理解植物生物学,也为植物育种等领域提供了新的研究方向,可以为将来在更多物种中广泛开展单细胞水平的研究提供参考.
Cell is the most basic structural unit of life,and its division,differentiation,and function are determined by gene expression.For a long time,the research on gene expression has been mainly analyzed at the whole tissue or organ level by conventional transcriptome sequencing(RNA-seq),but this method may lead to the equalization of different cells,and neglect the heterogeneity.In recent years,high-resolution single-cell transcriptome sequencing(scRNA-Seq)based on high throughput sequencing technology has been developed,which can isolate and sequence single cells from the same tissue sample in the same experiment.Single-cell transcriptome sequencing can accurately identify cell types and distinguish cell differentiation status according to the gene expression profiles,reveal changes in gene expression at single cell level in tissues and analyze tissue heterogeneity.Therefore scRNA-Seq can provide a new insight into the molecular mechanism of cell differentiation in organisms.The first step of single-cell sequencing is to separate single cells.A series of methods have been developed to separate single cells,including enzymatic hydrolysis,separation of single cell nucleus,laser capture microdissection,dilution,fluorescence-activated cell sorting(FACS),droplet microfluidic control and BD Rhapsody microhole capture,etc.Different separation approaches have advantages and disadvantages,which are analyzed in this review,and can be selected according to different experimental requirements and conditions.Compared with animal cells,the acquisition of a plant single cell needs to be achieved by the preparation of protoplasts.Enzymatic hydrolysis is one of the most commonly used methods to obtain protoplasts,but the process of enzymatic hydrolysis may have an obvious bias to the expression of related genes,and some plant protoplasts with larger diameter may lead to blocking of sequencing equipment,which limits the application of scRNA-Seq in plants.Despite this,the research of single-cell transcriptome in plants has increased
作者
安妍妍
张明惠
谭何新
Yanyan An;Minghui Zhang;Hexin Tan(School of Health Science and Engineering,University of Shanghai for Science and Technology,Shanghai 200093,China;Department Chinese Medicine Authentication,College of Pharmacy,Naval Medical University(Second Military Medical University),Shanghai 200433,China;Department of Pharmacy,Shanghai Fourth People’s Hospital Affiliated to Tongji University School of Medicine,Shanghai 200434,China)
出处
《科学通报》
EI
CAS
CSCD
北大核心
2024年第12期1586-1597,共12页
Chinese Science Bulletin
基金
国家优秀青年科学基金(82122072)
上海市自然科学基金(21ZR1477800)资助。
关键词
单细胞转录组学
单细胞分离
多组学
植物研究
single-cell transcriptomics sequencing
single cell isolation
multiple omics
plant research
