mDRIP-seq: a high-throughput method for R-loop mapping and quantitative assessment

Current methods for R-loop mapping need to perform DNA:RNA immunoprecipitation for each sample individually, with consequent limitations in throughput. Here, we develop and validate mDRIP-seq, a multi-sample barcoding and pooling method for R-loop mapping. We show mDRIP-seq performs equivalently as conventional methods, but with the merits of high throughput and cost-efficiency. We also show the simplicity of mDRIP-seq for relative and absolute quantitation of genomic R-loop fractions for multiple samples. Together, mDRIP-seq is a high-throughput and cost-efficient method for R-loop mapping and quantitative assessment and can be widely applied to large-scale dynamic profiles of these important structures for diverse organisms.当前用于R环（R-loop）定位的现有方法，需对每一份样本单独开展DNA:RNA免疫沉淀实验，由此带来通量层面的固有局限。本研究开发并验证了mDRIP-seq技术——一种可用于R环定位的多样本条码标记与混样富集方法。实验结果表明，mDRIP-seq的检测性能与传统方法相当，同时兼具高通量与高成本效益的显著优势。此外，该技术操作简便，可实现多份样本的基因组R环占比的相对定量与绝对定量分析。综上，mDRIP-seq是一种兼具高通量与成本效益优势的R环定位与定量评估技术，可广泛应用于多种生物中这类重要结构的大规模动态图谱分析。