RNA-seq of inner ear RNA from mice carrying a mutation in Zfp719 and wildtype controls at 3 different ages

The function of zinc finger protein 719 (Zfp719) remains largely unknown. The most closely related human gene ZNF175 encodes protein OTK18 which has 44% amino acid identity with mouse Zfp719, and is thought to be a transcriptional suppressor. Zfp719-deficient mice showed highly elevated auditory brainstem response (ABR) thresholds at 14 weeks in a high-throughput targeted mutagenesis programme (the Wellcome Trust Sanger Institute's Mouse Genetics Project). In order to understand the regulatory changes caused by the reduction in Zfp719, we carried out RNA-seq on RNA extracted from the inner ears of wildtypes and sex-matched homozygotes at three different ages; four days, two weeks and three weeks old, to look at expression before maturation of the auditory system, and before and after the onset of hearing loss. Strand-specific libraries were prepared using TruSeq stranded mRNA LT kit and sequenced on an Illumina NextSeq 500 machine as paired-end 150bp reads.锌指蛋白719（zinc finger protein 719，Zfp719）的功能目前仍鲜为人知。与其亲缘关系最近的人类同源基因ZNF175编码蛋白OTK18，该蛋白与小鼠Zfp719的氨基酸序列一致性达44%，被认为是一种转录抑制因子。在高通量靶向诱变项目——惠康信托桑格研究所小鼠遗传学计划（Wellcome Trust Sanger Institute's Mouse Genetics Project）中，Zfp719缺陷型小鼠在14周龄时的听觉脑干反应（auditory brainstem response, ABR）阈值显著升高。为探究Zfp719表达下调所引发的调控变化，我们针对三个不同年龄段（出生后4天、2周及3周）的野生型小鼠与性别匹配的纯合突变小鼠的内耳组织提取RNA，开展RNA测序实验，以期观测听觉系统成熟前、听力损失发病前后的基因表达水平。本研究采用TruSeq链特异性mRNA LT试剂盒构建链特异性测序文库，并在Illumina NextSeq 500测序平台上以150bp双端读长模式完成测序。