Transcription factor-like 5 is a potential DNA/RNA binding protein essential for maintaining male fertility in mice [RNA-seq]. Transcription factor-like 5 is a potential DNA/RNA binding protein essential for maintaining male fertility in mice [RNA-seq]

Tissue-specific transcriptional factors often play key roles in the development of specific cell lineages. Transcription factor-like 5 (TCFL5) is a testis-specific protein that contains the basic helix-loop-helix domain but the in vivo functions of TCFL5 remained unknown. Here, we generated CRISPR/Cas9-mediated knockout mice to dissect the function of TCFL5 in mouse testes. mRNA profiles of pachytene spermatocytes from Tcfl5+/+ and Tcfl5+/- mice sere generated by high throughput sequencing. The bound genes of TCFL5 in wild-type mouse testes were analyzed by ChIP-sequencing. Overall design: Pachytene spermatocytes were isolated from the testis of Tcfl5+/+ and Tcfl5+/- mice respectively. mRNA profiles of these cells were generated by high throughput sequencing, in two duplicate samples, using BGISEQ-500 platform of BGI Tech. Purified DNA by ChIP was also sequenced on the BGISEQ-500 platform of BGI Tech.

组织特异性转录因子通常在特定细胞谱系的发育进程中发挥关键调控作用。转录因子样5（TCFL5）是一种睾丸特异性蛋白质，含有碱性螺旋-环-螺旋（basic helix-loop-helix）结构域，但目前关于TCFL5的体内功能仍未明确。本研究构建了CRISPR/Cas9介导的基因敲除小鼠，以此解析TCFL5在小鼠睾丸中的功能。我们通过高通量测序获取了Tcfl5+/+与Tcfl5+/-小鼠粗线期精母细胞的mRNA表达谱；同时采用染色质免疫共沉淀测序（ChIP-sequencing）分析了野生型小鼠睾丸中TCFL5的结合靶基因。实验整体设计如下：分别从Tcfl5+/+与Tcfl5+/-小鼠的睾丸中分离粗线期精母细胞，设置两组生物学重复样本，采用华大基因（BGI Tech）的BGISEQ-500平台完成mRNA高通量测序；此外，通过染色质免疫共沉淀纯化得到的DNA同样使用BGI Tech的BGISEQ-500平台进行测序。