Cell-free gene expression (CFE) systems empower synthetic biologists to build biological molecules and processes outside of living intact cells. The foundational principle is that precise, complex biomolecular transformations can be conducted in purified enzyme or crude cell lysate systems. This concept circumvents mechanisms that have evolved to facilitate species survival, bypasses limitations on molecular transport across the cell wall, and provides a significant departure from traditional, cell-based processes that rely on microscopic cellular “reactors.” In addition, cell-free systems are inherently distributable through freeze-drying, which allows simple distribution before rehydration at the point-of-use. Furthermore, as cell-free systems are nonliving, they provide built-in safeguards for biocontainment without the constraints attendant on genetically modified organisms. These features have led to a significant increase in the development and use of CFE systems over the past two decades. Here, we discuss recent advances in CFE systems and highlight how they are transforming efforts to build cells, control genetic networks, and manufacture biobased products.

中文翻译：

---

游离基因表达：方法和应用


无细胞基因表达 （CFE） 系统使合成生物学家能够在完整活细胞之外构建生物分子和过程。基本原理是可以在纯化的酶或粗细胞裂解物系统中进行精确、复杂的生物分子转化。这个概念规避了已经进化为促进物种生存的机制，绕过了分子跨细胞壁运输的限制，并与依赖于微观细胞“反应器”的传统基于细胞的过程大相径庭。此外，无细胞系统本质上可以通过冷冻干燥进行分配，这允许在使用点再水化之前进行简单的分配。此外，由于无细胞系统是无生命的，它们为生物防护提供了内置的保护措施，而不受转基因生物的限制。在过去的二十年里，这些功能导致 CFE 系统的开发和使用显著增加。在这里，我们讨论了 CFE 系统的最新进展，并重点介绍了它们如何改变构建细胞、控制遗传网络和制造生物基产品的努力。