NIH Research Festival
During the process of spermatogenesis, significant epigenetic reprogramming occurs in the genome. CTCFL, a DNA-binding protein is expressed early in spermatogenesis and is widely recognized as a transcriptional regulator. Recent studies have suggested that CTCFL may have additional functions in organizing chromatin and controlling transposable elements, both of which are crucial for maintaining genome integrity. Our objective is to investigate the potential involvement of CTCFL in these processes. However, two major challenges have hindered progress in this area. Firstly, obtaining human germ cells that express CTCFL involves invasive procedures. Secondly, antibodies that target mouse CTCFL result in non-specific binding profiles in ChIP-seq analysis. Consequently, it is difficult to acquire human germ cells and mouse germ cells are unsuitable for analysis. To overcome these limitations, we have developed a novel humanized mouse model that expresses human CTCFL in mouse germ cells. This model enables us to utilize antibodies that have been verified for their specificity in targeting human CTCFL. By performing ChIP-seq analysis, we have discovered thousands of previously unidentified binding sites of CTCFL in germ cells. This breakthrough has set the foundation for our ongoing multi-omics approach, incorporating single-cell RNA-seq, ATAC-seq, and other NGS techniques to assess the epigenetic landscape in humanized germ cells and perform a comprehensive analysis of CTCFL functions during spermatogenesis.
Scientific Focus Area: Genetics and Genomics
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