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Functional Mutations in the microRNA-155 Promoter Modulate its Transcription Efficiency and Expression

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Abstract

Limited research has been conducted on porcine miR-155 promoters, and previous study from our group have identified two haplotypes (TT and CC) in different pig breeds, each associated with five fully linked mutation sites within or near the miR-155 gene (Li et al. Dev Comp Immunol 39(1):110–116, 2013). In this study, the promoter region of porcine miR-155 was screened, and two important transcription factors, Foxp3 and RelA, were identified. The binding ability of Foxp3 protein was found to be affected by the first mutation site (A/C) using EMSA analysis. In vitro experiments revealed that the expression level of miR-155 was significantly higher in the C haplotype compared to the T haplotype. Additionally, northern blotting assays indicated that both the first mutation site (A/C) and the fourth mutation site (G/T) had a significant impact on miR-155 expression levels. These findings provide further insights into the transcriptional regulation of porcine miR-155 and identify crucial mutation sites that influence miR-155 expression. This knowledge can serve as a basis for identifying potential molecular markers associated with disease resistance in swine.

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Data Availability

The authors declare that the main data supporting the findings of this study are available within the manuscript.

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Funding

This work was supported by the National Natural Science Foundation Youth Fund of China (Grant number 31702102).

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Authors and Affiliations

  1. College of Animal Science and Technology, Henan University of Animal Husbandry and Economy, No. 6 Longzi North Road, Zhengdong New District, Zhengzhou, 450046, Henan, China

    Congcong Li, Wanxia Zhao, Huijie Zhou, Jiao Wu, Yong Huo, Dongfeng Jiang, Qiuliang Xu & Wantao Li

  2. Henan Key Laboratory of Unconventional Feed Resources Innovative Utilization, Henan University of Animal Husbandry and Economy, Zhengzhou, Henan, China

    Xiangbo Ji & Kun Liu

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  1. Congcong Li
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Contributions

CL conceived and designed the study. CL and WZ processed the vector construction, while CL, YH, and WZ performed the cell culture, transfection, and luciferase assays. CL, JW, KL, and HZ performed the western blotting analysis, while EMSA. CL, DJ, XJ, and HZ performed northern blotting. CL, WL, and QX wrote the manuscript. All authors read and approved the final version of the manuscript.

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Correspondence to Congcong Li.

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12033_2023_857_MOESM1_ESM.rar

Supplementary figure1. No luciferase activity was detected in PK-15 cells transfected with pGL3-bicpro1 or pGL3-bicQ1/Q2/Q3 constructs. A pRL-TK vector that provided constitutive expression of Renilla luciferase was co-transfected as an internal control. The results are presented as mean±SEM (n=3). Supplementary table 1. Primers used for plasmid construction in this study. Supplementary table 2. Probes used for EMSA assay. Supplementary table 3. Probes used for Northern blot. (RAR 301 KB)

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Li, C., Zhao, W., Zhou, H. et al. Functional Mutations in the microRNA-155 Promoter Modulate its Transcription Efficiency and Expression. Mol Biotechnol (2023). https://doi.org/10.1007/s12033-023-00857-1

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