A wide range of sequencing methods has been developed to assess nascent RNA transcription and resolve the single-nucleotide position of RNA polymerase genome-wide. These techniques are often burdened with high input material requirements and lengthy protocols. We leveraged the template-switching properties of thermostable group II intron reverse transcriptase (TGIRT) and developed Butt-seq (bulk analysis of nascent transcript termini sequencing), which can produce libraries from purified nascent RNA in 6 h and from as few as 10,000 cells—an improvement of at least 10-fold over existing techniques. Butt-seq shows that inhibition of the superelongation complex (SEC) causes promoter-proximal pausing to move upstream in a fashion correlated with subnucleosomal fragments. To address transcriptional regulation in a tissue, Butt-seq was used to measure the circadian regulation of transcription from fly heads. All the results indicate that Butt-seq is a simple and powerful technique to analyze transcription at a high level of resolution.

中文翻译：

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Butt-seq：一种轻松分析转录的新方法

现已开发出多种测序方法来评估新生 RNA 转录并解析 RNA 聚合酶在全基因组范围内的单核苷酸位置。这些技术通常面临着高输入材料要求和冗长的协议的负担。我们利用热稳定 II 族内含子逆转录酶 (TGIRT) 的模板转换特性，开发了 Butt-seq（新生转录本末端测序的批量分析），它可以在 6 小时内从纯化的新生 RNA 中生成文库，并且仅需 10,000 个细胞——比现有技术至少提高 10 倍。Butt-seq 表明，抑制超延伸复合物 (SEC) 会导致启动子近端暂停以与亚核小体片段相关的方式向上游移动。为了解决组织中的转录调控问题，Butt-seq 用于测量果蝇头部转录的昼夜节律调节。所有结果都表明 Butt-seq 是一种简单而强大的技术，可以以高分辨率分析转录。