In vivo generation of DNA sequence diversity for cellular barcoding.
In vivo generation of DNA sequence diversity for cellular barcoding.
复制标题
体内生成用于细胞条形码的 DNA 序列多样性。
DOI：
10.1093/nar/gku604
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发表时间：
2014
影响因子：
14.9
通讯作者：
Zador,AnthonyM
中科院分区：
文献类型：
--
作者：
Peikon,IanD;Gizatullina,DianaI;Zador,AnthonyM
异质性是生物系统普遍存在的特征。对此类系统的完整理解需要一种唯一地识别和跟踪各个组件及其相互交互的方法。我们开发了一种新方法，可以通过 DNA 测序恢复的遗传“条形码”在体内对单个细胞进行独特标记。我们的方法是一个由基因条形码盒组成的双组分系统，其片段由 Rci（一种位点特异性 DNA 转化酶）进行改组。该系统具有高度可扩展性，有可能产生数十亿的理论多样性。我们证明了该技术在大肠杆菌中的可行性。目前，这种方法可用于通过各种瓶颈跟踪微生物种群的动态。这种方法的进步应该被证明有助于跟踪网络内细胞的相互作用和/或复杂生物样品内的异质性。
Heterogeneity is a ubiquitous feature of biological systems. A complete understanding of such systems requires a method for uniquely identifying and tracking individual components and their interactions with each other. We have developed a novel method of uniquely tagging individual cellsin vivowith a genetic ‘barcode’ that can be recovered by DNA sequencing. Our method is a two-component system comprised of a genetic barcode cassette whose fragments are shuffled byRci, a site-specific DNA invertase. The system is highly scalable, with the potential to generate theoretical diversities in the billions. We demonstrate the feasibility of this technique inEscherichia coli. Currently, this method could be employed to track the dynamics of populations of microbes through various bottlenecks. Advances of this method should prove useful in tracking interactions of cells within a network, and/or heterogeneity within complex biological samples.