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<p>γ-globin genes (<i>HBG1</i> and <i>HBG2</i>) are usually expressed during fetal life, and almost no expression after birth. Therefore, the reactivation of <i>HBG</i> is a key target for the treatment of hemoglobinopathy. <i>ZFP64</i> is a C2H2 type zinc finger transcription factor, which has been shown to play an important role in the maintenance of gene expression in mixed lineage leukemia, and other C2H2 type zinc finger transcription factors (such as <i>ZFP410</i> and <i>ZFP644</i>) have been shown to regulate the expression of fetal hemoglobin (HbF) in thalassemia. This study aims to investigate the effect of <i>ZFP64</i> on mRNA expression of <i>HBG</i>. We performed bioinformatics analyses using the Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and protein-protein interaction (PPI) networks to identify genes and transcription factors associated with <i>ZFP64. ZFP64</i> was knocked out in K562 and HUDEP-2 cell lines by CRISPR-Cas9 electroporation, and the transcription levels of <i>ZFP64</i>, <i>HBB</i> and <i>HBG</i> were analyzed. In undifferentiated and 7-day differentiated HUDEP-2 cells, knocking down <i>ZFP64</i> resulted in a 1.5-fold and 2.5-fold increase in <i>HBG</i> mRNA expression, respectively (<i>p</i> &lt; 0.05). These findings suggest that <i>ZFP64</i> is a potential regulator of <i>HBG</i> expression and warrants further investigation as a therapeutic target in hemoglobinopathies.</p>

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Investigate the Effect of ZFP64 on mRNA Expression of HBG Based on Bioinformatics and Experimental Validation

  • 期刊论文

  • 发表日期：

    2025年5月20日

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  • Siqi Huang,
  • Jianfeng Wu,
  • Yinghong Yang,
  • Mingming Zhu,
  • Lihao Chen,
  • Shunhan Zhang,
  • Yi Yang,
  • Xiaofang Sun,
  • Yingjun Xie

摘要

γ-globin genes (HBG1 and HBG2) are usually expressed during fetal life, and almost no expression after birth. Therefore, the reactivation of HBG is a key target for the treatment of hemoglobinopathy. ZFP64 is a C2H2 type zinc finger transcription factor, which has been shown to play an important role in the maintenance of gene expression in mixed lineage leukemia, and other C2H2 type zinc finger transcription factors (such as ZFP410 and ZFP644) have been shown to regulate the expression of fetal hemoglobin (HbF) in thalassemia. This study aims to investigate the effect of ZFP64 on mRNA expression of HBG. We performed bioinformatics analyses using the Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and protein-protein interaction (PPI) networks to identify genes and transcription factors associated with ZFP64. ZFP64 was knocked out in K562 and HUDEP-2 cell lines by CRISPR-Cas9 electroporation, and the transcription levels of ZFP64, HBB and HBG were analyzed. In undifferentiated and 7-day differentiated HUDEP-2 cells, knocking down ZFP64 resulted in a 1.5-fold and 2.5-fold increase in HBG mRNA expression, respectively (p < 0.05). These findings suggest that ZFP64 is a potential regulator of HBG expression and warrants further investigation as a therapeutic target in hemoglobinopathies.