单细胞基因组学分析技术前沿.pdfVIP

HEREDITAS (Beijing) 2011 1 , 33(1): 17―24 ISSN 0253-9772 综述 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. , CT 06520-8005 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。 单细胞基因组测序; 第二代测序技术; 功能基因组学; 转录组分析; 染色质免疫共沉淀; CpG 甲基化 图谱 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 Abstract: The technological progress of the genomics has transformed life science research. The main objectives of ge- nomics 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 lim- ited 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