Abstract

Zinc finger and BTB domain containing 1(Zbtb1) is a transcriptional suppressor protein, and a member of the mammalian Zbtb gene family. Previous studies have shown that Zbtb1 is essential for T-cell development. However, the role of Zbtb1 in T-cell lymphoma is undetermined. In this study, an EL4 cell line with Zbtb1 deletion was constructed using the CRISPR-Cas9 technique. The expression profiles of microRNA and circRNA produced by the control and gene deletion groups were determined by RNA-seq. In general, 24 differentially expressed microRNA and 16 differentially expressed circRNA were found between normal group and gene deletion group. Through further analysis of differentially expressed genes, GO term histogram and KEGG scatter plot were drawn, and three pairs of miRNA and circRNA regulatory relationships were found. This study describes the differentially expressed microRNA and circRNA in normal and Zbtb1-deficient EL4 cell lines, thus providing potential targets for drug development and clinical treatment of T-cell lymphoma.

Highlights

  • Expression of Zbtb1 is necessary for normal lymphoid development

  • Constructing EL4 Cells With Zbtb1 Gene Knockout sgRNA Screening In Vitro The target fragment of the EL4 cell genome was amplified by PCR (Zbfb1_survy_F: ACGACATCTACTTCCAAGCACACA; Zbfb1_survy_R: GGTAAGCTCTTGTCGTGTTTTGGT), and the length of the fragment was 664 bp (Figure 1A), the target sequence was consistent with the National Center for Biotechnology Information (NCBI) sequence and the alignment results (Figure 1B)

  • The results showed that the subcutaneous growth rate of EL4 cells with Zbtb1 gene knockout in C57BL/6 mice was significantly slower than that of wild type EL4 cells

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Summary

Introduction

Zbtb maintains genomic integrity in immune progenitors during the process of replication and differentiation, while Zbtb deficiency increases DNA damage and p53-mediated apoptosis, mRNA encoding Zbtb is most highly expressed in hematopoietic stem cells, thymocytes and pre-B cells, In addition to its role in T cell development, it was demonstrated to be involved in the differentiation of B cells and NK cells (Siggs et al, 2012; Punwani et al, 2012; Cao et al, 2016; Cao et al, 2018). MiR-23b-3p inhibits breast cancer cell proliferation and tumor growth by targeting Zbtb to regulate aerobic glycolysis in tamoxifenresistant cells (Kim et al, 2014)

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