单细胞基因组学分析的技术前沿

doi:10.3724/SP.J.1005.2011.00017

遗传 ›› 2011, Vol. 33 ›› Issue (1): 17-24.doi: 10.3724/SP.J.1005.2011.00017

单细胞基因组学分析的技术前沿

潘星华, 朱海英, MARJANI Sadie L.

耶鲁大学医学院遗传系, 美国康耐狄克州纽黑文市 CT 06520-8005

收稿日期:2010-08-23 修回日期:2010-10-31 出版日期:2011-01-20 发布日期:2011-01-20
通讯作者: 潘星华 E-mail:xinghua.pan@yale.edu
基金资助:
单细胞转录组和甲基化组分析, NIH项目(编号: 1R21HD066457-01)资助

Technological advances in single-cell genomic analyses

PAN Xing-Hua, ZHU Hai-Ying, MARJANI Sadie L.

Department of Genetics, Yale University School of Medicine, New Haven CT 06520-8005, USA

Received:2010-08-23 Revised:2010-10-31 Online:2011-01-20 Published:2011-01-20
Contact: PAN Xin-hua E-mail:xinghua.pan@yale.edu

摘要/Abstract

摘要： 基因组学已经深刻地改变了生命科学的诸多领域的面貌。目前它的主要内容是新的全基因组碱基序列的测定和在全基因组范围内鉴定那些在不同水平上影响生命活动的基因群的功能和相互作用。为达此要求, 近年出现的第二代测序(深度测序)技术和基因芯片技术发挥了关键作用, 但是两者都需要足够的高质量的核酸样品。所以, 在只有或只能用单细胞或极少量细胞的情况下, 如果没有特殊手段, 上述分析往往不能常规、方便地进行。文章以DNA扩增为主线, 综合阐述了目前在单细胞(特别是微生物)全基因组测序和大基因组的靶向重测序, 以及对单细胞或微量细胞进行的基于深度测序或芯片杂交的功能基因组分析, 如转录组、ChIP和DNA的CpG甲基化分析等的最新策略和技术, 评价了单细胞基因组测序和功能基因组学各技术的特点并对发展前景进行了展望。

关键词: CpG甲基化图谱, 染色质免疫共沉淀, 单细胞基因组测序, 转录组分析, 功能基因组学, 第二代测序技术

Abstract: The technological progress of the genomics has transformed life science research. The main objectives of genomics are sequencing of new genomes and genome-wide identification of the function and the interaction of genes and their products. The recently developed second generation or next generation sequencing platforms and DNA microarray technology are immensely important and powerful tools for functional genomic analyses. However, their application is limited by the requirement of sufficient amounts of high quality nucleic acid samples. Therefore, when only a single cell or a very small number of cells are available or are preferred, the whole genomic sequencing or functional genomic objectives cannot be achieved conventionally and require a robust amplification method. This review highlights DNA amplification technologies and summarizes the strategies currently utilized for whole genome sequencing of a single cell, with specific focus on studies investigating microorganisms; An outline for targeted re-sequencing enabling the analysis of larger genomes is also provided. Furthermore, the review presents the emerging functional genomic applications using next-generation sequencing or microarray analysis to examine genome-wide transcriptional profile, chromatin modification and other types of protein-DNA binding profile, and CpG methylation mapping in a single cell or a very low quantity of cells. The nature of these technologies and their prospects are also addressed.

Key words: single-cell sequencing, next generation sequencing, functional genomics, transcriptome profiling, chromatin immunoprecipitation, CpG methylation mapping

潘星华，朱海英，MARJANI Sadie L.. 单细胞基因组学分析的技术前沿[J]. 遗传, 2011, 33(1): 17-24.

BO Xing-Hua, SHU Hai-Yang, MARJANI Sadie L.. Technological advances in single-cell genomic analyses[J]. HEREDITAS, 2011, 33(1): 17-24.

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