深度突变扫描技术在蛋白研究中的应用

Application of deep mutational scanning technology in protein research

摘要:

作为细胞结构与功能的中心参与者，蛋白质一直是生命科学研究的中心主题。分析蛋白质序列变异对其结构、功能的影响，是研究蛋白的重要手段之一。近年一种称为深度突变扫描(deep mutational scanning, DMS)的技术被广泛应用于蛋白研究领域，其通过高丰度DNA文库在蛋白特定区域平行引入成千上万种突变，经筛选后，利用高通量测序为每一种突变打分，从而揭示序列与功能之间的相关性。深度突变扫描以其高通量、快速简易、节省人工等特点，已经成为蛋白质功能研究以及蛋白工程改造的一种重要方法，目前已在蛋白进化、抗体改造、致病突变鉴定等蛋白研究的多个领域广泛应用。本综述简要概括了深度突变扫描技术的原理，重点介绍了其在哺乳动物细胞中的应用，同时分析了目前的技术瓶颈，旨在为相关研究提供参考。

Abstract:

As central players in cellular structure and function, proteins have long been central themes in life science research. Analyzing the impact of protein sequence variation on its structure and function is one of the important means to study proteins. In recent years, a technology called deep mutational scanning (DMS) has been widely used in the field of protein research. It introduces thousands of mutations in parallel in specific regions of proteins through high-abundance DNA libraries. After screening, high-throughput sequencing is employed to score each mutation, revealing sequence-function correlations. Due to its high-throughput, fast and easy, and labor-saving features, DMS has become an important method for protein function research and protein engineering. This review briefly summarizes the principle of DMS technology, highlighting its applications in mammalian cells. Moreover, this review analyzes the current technical bottlenecks, aiming to facilitate relevant research.