Department of Molecular & Medical Genetics, Knight Cancer Institute, Knight Cardiovascular Institute, Cancer Early Detection Advanced Research Center, Oregon Health and Science University, Portland, Oregon 97239, USA
Genome Res. 2021 Oct;31(10):1693-1705. doi: 10.1101/gr.275223.121.
It has been just over 10 years since the initial description of transposase-based methods to prepare high-throughput sequencing libraries, or "tagmentation," in which a hyperactive transposase is used to simultaneously fragment target DNA and append universal adapter sequences. Tagmentation effectively replaced a series of processing steps in traditional workflows with one single reaction. It is the simplicity, coupled with the high efficiency of tagmentation, that has made it a favored means of sequencing library construction and fueled a diverse range of adaptations to assay a variety of molecular properties. In recent years, this has been centered in the single-cell space with a catalog of tagmentation-based assays that have been developed, covering a substantial swath of the regulatory landscape. To date, there have been a number of excellent reviews on single-cell technologies structured around the molecular properties that can be profiled. This review is instead framed around the central components and properties of tagmentation and how they have enabled the development of innovative molecular tools to probe the regulatory landscape of single cells. Furthermore, the granular specifics on cell throughput or richness of data provided by the extensive list of individual technologies are not discussed. Such metrics are rapidly changing and highly sample specific and are better left to studies that directly compare technologies for assays against one another in a rigorously controlled framework. The hope for this review is that, in laying out the diversity of molecular techniques at each stage of these assay platforms, new ideas may arise for others to pursue that will further advance the field of single-cell genomics.
自最初描述基于转座酶的高通量测序文库制备方法（即“标签化”）以来，已经过去了 10 多年，在这种方法中，使用超活性转座酶同时对靶 DNA 进行片段化并添加通用接头序列。标签化有效地用一个单一的反应替代了传统工作流程中的一系列处理步骤。正是这种简单性，加上标签化的高效率，使其成为测序文库构建的首选方法，并激发了对各种分子特性进行多样化检测的各种适应性。近年来，这一方法主要集中在单细胞领域，开发了一系列基于标签化的检测方法，涵盖了调控景观的大部分区域。迄今为止，已经有许多关于可以进行分析的分子特性的单细胞技术的优秀综述。这篇综述则是围绕标签化的核心组件和特性以及它们如何使创新的分子工具得以开发，以探测单细胞的调控景观展开的。此外，关于单个技术提供的细胞通量或数据丰富度的详细信息并未讨论。这些指标变化迅速且高度依赖于样本，最好留待在严格控制的框架下直接比较技术对检测的研究。本文希望通过展示这些检测平台各个阶段的分子技术多样性，为其他人提供新的思路，从而进一步推动单细胞基因组学领域的发展。
