中国畜牧兽医 ›› 2023, Vol. 50 ›› Issue (2): 479-489.doi: 10.16431/j.cnki.1671-7236.2023.02.006

• 生物技术 • 上一篇    下一篇

单细胞转录组测序技术及其在肌内脂肪细胞研究中的应用

张久盘1, 马青1, 王锦1, 马丽娜1, 魏大为2, 梁小军1   

  1. 1. 宁夏农林科学院动物科学研究所, 银川 750002;
    2. 宁夏大学农学院, 银川 750021
  • 收稿日期:2022-06-24 出版日期:2023-02-05 发布日期:2023-02-06
  • 通讯作者: 梁小军 E-mail:Lxj0520@163.com
  • 作者简介:张久盘,E-mail:zhangjiupan@163.com
  • 基金资助:
    宁夏自然科学基金项目(2021AAC05007、2021A0491);国家自然科学基金项目(32060744、32202641);宁夏自治区滩羊育种专项(2018NYYZ0401);国家现代农业(肉牛牦牛)技术体系资助(CARS-37)

Single-cell RNA Sequencing Technology and Its Application in the Study of Intramuscular Adipocytes

ZHANG Jiupan1, MA Qing1, WANG Jin1, MA Lina1, WEI Dawei2, LIANG Xiaojun1   

  1. 1. Institute of Animal Science, Ningxia Academy of Agriculture and Forestry Sciences, Yinchuan 750002;
    2. School of Agriculture, Ningxia University, Yinchuan 750021
  • Received:2022-06-24 Online:2023-02-05 Published:2023-02-06

摘要: 单细胞转录组测序(single-cell RNA sequencing,scRNA-seq)技术在单细胞水平对转录组进行测序,可从表达量、细胞量及细胞组成等多种角度描述样本,主要包括单细胞分离、mRNA反转录、文库构建、转录组测序和数据分析等步骤。单细胞分离是scRNA-seq技术操作的第一步,常用的细胞分离方法有连续稀释法、显微操作法、荧光激活流式细胞术、激光捕获显微切割术和微流控技术。基于不同细胞捕获、cDNA扩增和文库构建开发了许多scRNA-seq方法,如CEL-seq2、Drop-seq、MARS-seq、SCRB-seq、Smart-seq、Smart-seq2等。与传统测序方法相比,scRNA-seq可识别单个细胞的基因表达信息、记录细胞的空间位置、跟踪不同细胞谱系在分化过程中的轨迹,这为研究难以大量提取的肌内脂肪细胞分子特异性和发生分化过程带来极大便利。作者简要介绍了scRNA-seq技术的关键步骤,比较分析了单细胞分离和转录组测序方法的优缺点,综述了肌内脂肪细胞的起源以及scRNA-seq在肌内脂肪细胞的亚群和标记基因鉴定以及细胞分化轨迹追踪中的应用,以期为肌内脂肪细胞的深入研究提供参考依据。

关键词: 单细胞转录组测序; 肌内脂肪细胞; 细胞异质性

Abstract: Single-cell RNA sequencing (scRNA-seq) technology is used to sequence the transcriptome at the single cell level,which can describe samples from multiple perspectives such as expression amount,cell size and cell composition,mainly including single cell isolation,mRNA reverse transcription,library construction,transcriptome sequencing and data analysis.Single cell isolation is the first step of this technology. Common cell isolation methods include serial dilution,micromanipulation,fluorescent activated cell sorting,laser capture microdissection and microfluidics.Many transcriptome sequencing methods based on different cell capture,cDNA amplification and library construction have been developed,such as CEL-seq2,Drop-seq,MARS-seq,SCRB-seq,Smart-seq,Smart-seq2,etc.Compared with the traditional sequencing method,scRNA-seq can identify the gene expression information of a single cell,record the spatial location of the cell,and track the trajectory of different cell lineages in the differentiation process,which brings great convenience for studying the molecular specificity of intramuscular adipocytes that are difficult to extract in large quantities and the occurrence and differentiation process.The author briefly introduced the key steps of scRNA-seq technology,compared and analyzed the advantages and disadvantages of single cell isolation and transcriptome sequencing methods,reviewed the origin of intramuscular adipocytes,and the application of scRNA-seq in the identification of subgroups and marker genes of intramuscular adipocytes,as well as the tracking of cell differentiation trajectory,in order to provide a reference for the further study of intramuscular adipocytes.

Key words: single-cell RNA sequencing; intramuscular adipocytes; cell heterogeneity

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