Cross-linking and immunoprecipitation (CLIP) analysis for identifying binding sites of Tet2 in bone marrow derived mast cell

To transcriptome-widely identify the mRNA targets of Tet2 and investigate the immunological function of Tet2 at RNA regulation level in vivo, we performed CLIP-seq of Tet2 in BMMCs. We performed three bio-replicates of CLIP-seq for getting potential Tet2-binding mRNAs.Using data of normalized tag numbers in the common peaks, we found that biological replicates correlated well with each other.More than 60% of the peaks located in genic regions, which were preferentially enriched in coding sequencing (CDS) among mature mRNA elements. Overall design: Systematically identify Tet2-binding RNAs in biological triplicate.

为在全转录组范围内鉴定Tet2的mRNA靶标，并在体内RNA调控维度探究Tet2的免疫学功能，我们在骨髓源性肥大细胞（BMMCs）中开展了Tet2的紫外交联免疫沉淀测序（CLIP-seq）实验。为获取潜在的Tet2结合mRNA，我们设置了三次生物学重复的CLIP-seq实验。通过分析共同峰中的标准化标签计数数据，我们发现各生物学重复之间具有良好的相关性。超过60%的峰定位于基因区域，在成熟mRNA元件中，此类峰优先富集于编码序列（CDS）。实验整体设计：通过三次生物学重复系统鉴定Tet2结合的RNA。