单细胞测序技术及其在毛囊发育中的应用

doi:10.13560/j.cnki.biotech.bull.1985.2020-1491

摘要/Abstract

摘要：

毛囊是一种具有独特结构的、呈周期性生长的微型器官,其形态发生始于胚胎期,由表皮上皮、间质真皮及特殊衍生物经过一系列相互作用诱导形成。出生后个体的毛囊开始进行周期性循环,包括生长期、退行期和休止期。单细胞转录组测序（single-cell RNA sequence,scRNA-seq）是一种新的测序方法,通过制备单细胞悬液或细胞群,利用二代测序（next generation sequencing）来识别单个细胞的基因表达信息,主要用于分析细胞间遗传和基因表达水平的差异,更好地了解单个细胞在微环境中的具体作用。通过单细胞转录组测序可以揭示复杂和稀有细胞群体以及基因之间的调节关系,跟踪不同细胞谱系在发育过程中的轨迹。本文就单细胞转录组测序技术及其在毛囊发育调控研究中的应用展开叙述,以期为揭示毛囊发育的分子调控机制研究提供理论参考。

关键词: 单细胞转录组测序, 单细胞分离技术, 毛囊发育

Abstract:

Hair follicle is a kind of micro-organ with unique structure and periodic growth,its morphogenesis begins in the embryonic stage and is induced by a series of interactions among epidermis epithelium,interstitial dermis and special derivatives. After birth,individual hair follicles begin to cycle periodically,including growth period,retrogression period and resting period. Single-cell RNA sequence（scRNA-seq）is a new sequencing method,mainly through the preparation of single cell suspension or cell groups and using the next generation sequencing to identify the gene expression information of the single cell. It is mainly used to analyze the differences of genetic and gene expression levels among cells,to better understand the specific role of a single cell in the microenvironment. The regulatory relationship between complex and rare cell populations and genes may be revealed via scRNA-seq,and the developmental trajectories of different cell lineages can be traced. This paper describes the scRNA-seq and its application in the regulation of hair follicle development,aiming to provide theoretical reference for revealing the molecular regulation mechanism of hair follicle development.

Key words: single-cell RNA sequencing, single cell isolation technique, hair follicle development

叶娜, 张晓兰, 包鹏甲, 王兴东, 阎萍, 潘和平. 单细胞测序技术及其在毛囊发育中的应用[J]. 生物技术通报, 2021, 37(10): 245-256.

YE Na, ZHANG Xiao-lan, BAO Peng-jia, WANG Xing-dong, YAN Ping, PAN He-ping. Single Cell Sequencing Technology and Its Application in Hair Follicle Development[J]. Biotechnology Bulletin, 2021, 37(10): 245-256.

图/表 2

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方法 Method	cDNA合成 cDNA synthesis	扩增方法 Amplification method	捕获细胞量 Quantity of captured cells	cDNA覆盖 cDNA cover	Barcode	UMI	参考文献Reference
Tang2009	ployA尾	PCR	~10	3'端偏移的全长	No	No	[23]
Quartz-seq	ployA尾	PCR	1 000-10 000	3'端偏移的全长	No	No	[76]
SUPeR-seq	ployA尾	PCR	~10	全长	Yes	No	[67]
MATQ-seq	ployA尾	PCR	10-100	全长	Yes	Yes	[68]
STRT-seq	5'端模板转换	PCR	100-1 000	5'末端	Yes	Yes	[37]
Smart-seq	5'端模板转换	PCR	100-1 000	3'偏倚的全长	No	No	[41]
10xGenomics	5'端模板转换	PCR	1 000-10 000	3'末端	Yes	Yes	[44]
Seq-well	5'端模板转换	PCR	100-1 000	3'末端	Yes	Yes	[46]
Microwell-seq	5'端模板转换	PCR	100-10 000	全长	Yes	Yes	[45]
Drop-seq	5'端模板转换	PCR	1 000-10 000	3'末端	Yes	Yes	[43]
CEL-seq	IVT	IVT	100-1 000	3'末端	Yes	Yes	[73]
MARS-seq	IVT	IVT	1 000-5 000	3'末端	Yes	Yes	[27]
inDrops	IVT	IVT	1 000-10 000	3'末端	Yes	Yes	[42]

方法 Method	cDNA合成 cDNA synthesis	扩增方法 Amplification method	捕获细胞量 Quantity of captured cells	cDNA覆盖 cDNA cover	Barcode	UMI	参考文献Reference
Tang2009	ployA尾	PCR	~10	3'端偏移的全长	No	No	[23]
Quartz-seq	ployA尾	PCR	1 000-10 000	3'端偏移的全长	No	No	[76]
SUPeR-seq	ployA尾	PCR	~10	全长	Yes	No	[67]
MATQ-seq	ployA尾	PCR	10-100	全长	Yes	Yes	[68]
STRT-seq	5'端模板转换	PCR	100-1 000	5'末端	Yes	Yes	[37]
Smart-seq	5'端模板转换	PCR	100-1 000	3'偏倚的全长	No	No	[41]
10xGenomics	5'端模板转换	PCR	1 000-10 000	3'末端	Yes	Yes	[44]
Seq-well	5'端模板转换	PCR	100-1 000	3'末端	Yes	Yes	[46]
Microwell-seq	5'端模板转换	PCR	100-10 000	全长	Yes	Yes	[45]
Drop-seq	5'端模板转换	PCR	1 000-10 000	3'末端	Yes	Yes	[43]
CEL-seq	IVT	IVT	100-1 000	3'末端	Yes	Yes	[73]
MARS-seq	IVT	IVT	1 000-5 000	3'末端	Yes	Yes	[27]
inDrops	IVT	IVT	1 000-10 000	3'末端	Yes	Yes	[42]