Institute of Cellular and Molecule Biology, University of Texas at Austin, TX 78712, USA.
Methods. 2013 Mar 15;60(1):75-80. doi: 10.1016/j.ymeth.2012.03.008. Epub 2012 Apr 3.
In vitro compartmentalization (IVC) is a method to generate numerous, small, aqueous compartments (up to 10(10) compartments per ml) by mixing water, surfactants, and oil. The water phase is surrounded by surfactants and an oil phase, and to a first approximation each water-in-oil compartment is like an artificial cell. By introducing single genes into compartments that are competent for transcription and translation, these cell-like compartments can synthesize RNA protein variants in libraries. Screening or selecting for function has in turn led to schemes for the directed evolution of biomolecules. However, IVC selections can cover larger library sizes, and provide greater control over selection conditions and stringencies. The key issue in designing and executing IVC selections is how to couple genotype and phenotype, and in this review we have organized and presented a variety of mechanisms by which proteins and RNA can attach to or amplify their own templates following emulsification and selection.
体外分隔（IVC）是一种通过混合水、表面活性剂和油来生成大量小的水相隔间（每毫升可达 10^10 个隔间）的方法。水相被表面活性剂和油相包围，并且在第一近似下，每个油包水隔间都像一个人工细胞。通过将单个基因引入具有转录和翻译能力的隔间中，可以在这些类细胞隔间中合成 RNA 蛋白变体文库。筛选或选择功能反过来又导致了生物分子的定向进化方案。然而，IVC 选择可以覆盖更大的文库大小，并提供对选择条件和严格程度的更大控制。在设计和执行 IVC 选择时的关键问题是如何将基因型和表型联系起来，在这篇综述中，我们组织并介绍了多种机制，通过这些机制，蛋白质和 RNA 可以在乳化和选择后附着或扩增它们自己的模板。
