Griffiths Andrew D, Tawfik Dan S
Institut de Science et d'Ingénierie Supramoléculaires (ISIS), 8 allée Gaspard Monge, BP 70028, F-67083 Strasbourg Cedex, France.
Trends Biotechnol. 2006 Sep;24(9):395-402. doi: 10.1016/j.tibtech.2006.06.009. Epub 2006 Jul 14.
Biochemical and genetic assays can be both miniaturized and parallelized by compartmentalization in living cells. In vitro compartmentalization (IVC) offers an alternative strategy based on partitioning reactions in water droplets dispersed to form microscopic compartments in water-in-oil emulsions. The cell-like volumes of these compartments (as low as one femtolitre), the ability to freely determine and regulate their content and the large number of compartments (>10(10) per millilitre emulsion) have provided the basis for a range of new, ultra-high-throughput, cell-free technologies. This review describes the scope and potential of IVC in areas such as in vitro evolution of proteins and RNAs, cell-free cloning and sequencing, genetics, genomics, and proteomics.
通过在活细胞中进行区室化,生化和遗传分析可以实现小型化和平行化。体外区室化(IVC)提供了一种替代策略,该策略基于在油包水乳液中分散形成微观区室的水滴中进行分区反应。这些区室的细胞样体积(低至一飞升)、自由确定和调节其内容物的能力以及大量的区室(每毫升乳液>10^10个)为一系列新的超高通量无细胞技术提供了基础。本综述描述了IVC在蛋白质和RNA的体外进化、无细胞克隆和测序、遗传学、基因组学和蛋白质组学等领域的范围和潜力。