Department of Parasitology, Guangxi Medical University, Nanning 530021, China.
Department of Cell Biology and Genetics, Guangxi Medical University, Nanning 530021, China.
Yi Chuan. 2024 Aug;46(8):589-602. doi: 10.16288/j.yczz.24-149.
Self-transcribing active regulatory region sequencing (STARR-seq) is a high-throughput sequencing method capable of simultaneously discovering and validating all enhancers within the genome. In this method, candidate sequences are inserted into plasmid vectors and electroporated into cells. Acting as both enhancers and target genes, the self-transcription of these sequences will also be enhanced by themselves. By sequencing the transcriptome and comparing the results with the non-inserted control, the locations and activity of enhancers can be determined. In traditional enhancer discovery strategies, the chromatin open regions and transcription active regions were sequenced and predicted as enhancers. However, the activity of these putative enhancers could only be validated one by one without a high-throughput method. STARR-seq solved this limitation, allowing simultaneous enhancers discovery and activity validation in a high-throughput manner. Since the introduction of STARR-seq, it has been widely used to discover enhancers and validate enhancer activity in a number of organisms and cells. In this review, we present the traditional enhancer prediction methods and the basic principles, development history, specific applications of STARR-seq, and its future prospects, aiming to provide a reference for researchers in related fields conducting enhancer studies.
自我转录活性调控区测序（STARR-seq）是一种高通量测序方法，能够同时发现和验证基因组内的所有增强子。在这种方法中，候选序列被插入质粒载体并电穿孔到细胞中。这些序列作为增强子和靶基因的双重作用，其自身的自我转录也将被增强。通过对转录组进行测序，并将结果与未插入的对照进行比较，可以确定增强子的位置和活性。在传统的增强子发现策略中，染色质开放区域和转录活跃区域被测序并预测为增强子。然而，这些假定的增强子的活性只能逐个进行验证，而没有高通量的方法。STARR-seq 解决了这一局限性，能够以高通量的方式同时发现增强子并验证其活性。自 STARR-seq 问世以来，它已被广泛用于在多种生物体和细胞中发现增强子并验证增强子活性。在这篇综述中，我们介绍了传统的增强子预测方法以及 STARR-seq 的基本原理、发展历史、具体应用及其未来前景，旨在为从事增强子研究的相关领域的研究人员提供参考。
