Genome-wide R-loops dynamics and its alteration by Nkapl knockout in mouse testes with ssDRIP-seq

To investigate genome-wide R-loops during meiosis exit and R-loop profiles changes with Nkapl knockout, we employed ssDRIP-seq (single-strand DNA ligation-based library preparation after DNA:RNA hybrid immunoprecipitation by S9.6 and sequencing) in wild-type and Nkapl-KO testes at P21. For ssDRIP-seq, we have biological duplicates. Input was separated, and S9.6 antibody was used for immunoprecipitation of DNA:RNA hybrid in 21d wild-type and Nkapl KO mouse testes.

为探究减数分裂退出阶段的全基因组R环（R-loop）分布特征，以及Nkapl基因敲除（knockout, KO）对R环图谱的调控变化，我们对21日龄（P21）的野生型与Nkapl基因敲除型小鼠睾丸样本，采用了ssDRIP-seq技术：该技术的具体流程为，通过S9.6抗体完成DNA:RNA杂合物的免疫沉淀后，基于单链DNA连接法构建测序文库并进行测序。本实验中ssDRIP-seq设置了生物学重复样本；我们同时分离得到Input对照样本，并使用S9.6抗体对21日龄野生型及Nkapl基因敲除型小鼠睾丸中的DNA:RNA杂合物进行免疫沉淀。