Hemoglobin is a vital component of the circulatory system in higher organisms, and its dysfunction is closely associated with conditions such as anemia and cardiovascular diseases. Antarctic icefish, whose blood appears white due to the absence of functional hemoglobin, serve as a unique model for studying the biological functions of hemoglobin. We identified a conserved non-coding element （CNE） located in the intergenic region chr6：43629743-43629779, upstream of the foxp1b gene, which may play a key role in hemoglobin production in fish. However, its specific regulatory function remains unclear. In this study, we utilized CRISPR/Cas9 gene-editing technology to generate zebrafish heterozygous mutants with knockouts of this CNE. Hemoglobin staining was subsequently performed to compare hemoglobin production between the wild-type control group and the mutant zebrafish. The results demonstrated that the knockout of chr6：43629743-43629779 significantly reduced hemoglobin production in zebrafish. Additionally, qRT-PCR（Quantitative Reverse Transcription Polymerase Chain Reaction） analysis revealed that foxp1b gene expression was significantly lower in the mutant zebrafish compared to the wild-type controls. This study indicates the molecular mechanism by which CNEs regulate hemoglobin production by modulating the expression of downstream genes. It highlights the critical role of conserved non-coding elements in hemoglobin synthesis and offers new insights into the regulation of this essential protein.