Nat Biotechnol:开发出进行单一细胞全基因组测序的新技术

 

    近日,刊登在国际杂志Nature Biotechnology上的一篇研究论文中,来自加州大学的研究者通过研究,成功对人类大脑单一神经元细胞和大肠杆菌单一细胞进行了全基因组的测序,研究者仅需要用12纳升的流动液体就可以实现对单一细胞的全基因组测序。
 
    文中,Kun Zhang教授表示,我们的前期试验显示,来自同一大脑的单一神经元具有不同的遗传组成部分,我们的研究可以使得众多科学家比较出单一细胞间更加细微的遗传差异情况;对单一细胞进行测序是很多领域科学家长期以来比较感兴趣的一个研究领域,比如,探索单一细胞的遗传组成部分将帮助研究者鉴别出很多不能够在实验室条件下培养的细菌。
 
    当然对单一细胞的研究也将用于用于揭示癌细胞的发生机制,目前研究者仅仅需要12纳升的微流体就可以实现对单一细胞的全基因组进行测序。研究者Jeff Gole解释道,通过降低扩增反应的用量,我们就可以增加DNA模板的浓度,从而可以帮助改善扩增的完整性以及降低DNA的污染。
 

    相比此前对大肠杆菌单一细胞的测序来讲,这种新型技术可以更加有效地对大肠杆菌的单一细胞进行测序。最后研究者Zhang表示,我们对我们的研究结果非常激动,我们希望未来全世界的研究者将使用这项新型技术来实现对单一细胞的全基因组测序。

 

doi:10.1038/nbt.2720

 

 Massively parallel polymerase cloning and genome sequencing of single cells using nanoliter microwells

 

  Jeff Gole, Athurva Gore,        Andrew Richards, Yu-Jui Chiu,    Ho-Lim Fung, Diane Bushman, Hsin-I Chiang,     Jerold Chun, Yu-Hwa Lo        & Kun Zhang

 

  Genome sequencing of single cells has a variety of applications, including characterizing difficult-to-culture microorganisms and identifying somatic mutations in single cells from mammalian tissues. A major hurdle in this process is the bias in amplifying the genetic material from a single cell, a procedure known as polymerase cloning. Here we describe the microwell displacement amplification system (MIDAS), a massively parallel polymerase cloning method in which single cells are randomly distributed into hundreds to thousands of nanoliter wells and their genetic material is simultaneously amplified for shotgun sequencing. MIDAS reduces amplification bias because polymerase cloning occurs in physically separated, nanoliter-scale reactors, facilitating the de novo assembly of near-complete microbial genomes from single Escherichia coli cells. In addition, MIDAS allowed us to detect single-copy number changes in primary human adult neurons at 1- to 2-Mb resolution. MIDAS can potentially further the characterization of genomic diversity in many heterogeneous cell populations.