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CRISPR/Cas9-Mediated Disruption of ZNF543 Gene: An Approach Toward Discovering Its Relation to TRIM28 Gene in Parkinson’s Disease

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Abstract

Genetic studies of familial forms of Parkinson’s disease (PD) have shown that the ZNF543 gene is a candidate gene that operates relevant to this disease. However, until now, there is no evidence for ZNF543 gene function in PD, and mechanisms resulting from its mutation have not been elucidated. Given the same genetic location of the ZNF543 gene with TRIM28 and their effects on PD pathogenesis, we surmised that ZNF543 might act as a transcription factor for TRIM28 gene expression. By knocking out the ZNF543 gene via the CRISPR/Cas9 editing platform, we assessed the functional effect of loss of expression of this gene on TRIM28 gene expression. Four sgRNAs with different PAM sequences were designed against two parts of the regulatory region of ZNF543 gene, and highly efficient disruption of ZNF543 expression in human neuroblastoma cell line was evaluated by polymerase chain reaction and T7 endonuclease assay. Moreover, evaluation of TRIM28 gene expression in ZNF543-knocked-out cells indicated a significant increase in TRIM28 gene expression, suggesting that ZNF543 probably regulates the expression of TRIM28. This approach offers a window into pinpointing the mechanism by which ZNF543 gene mutations mediate PD pathogenicity.

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Data Availability

The data sets generated and analyzed in the current study are available from the corresponding author on reasonable request.

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Acknowledgements

The authors express appreciation to the Shahid Bahonar University of Kerman for its support of this investigation.

Funding

Not Applicable.

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Authors and Affiliations

  1. Department of Genetic, Faculty of Biological Science, Tarbiat Modares University, Tehran, Iran

    Mohammad Hashemabadi

  2. Department of Biology, Faculty of Science, Shahid Bahonar University of Kerman, Kerman, Iran

    Hosseinali Sasan, Saeed Esmaeili-Mahani & Hadi Ravan

  3. Department of Biochemistry, Faculty of Biological Science, Tarbiat Modares University, Tehran, Iran

    Mojdeh Amandadi

  4. Chair of Crop Science and Plant Biology, Institute of Agricultural and Environmental Sciences, Estonian University of Life Sciences, Fr. R.Kreutzwaldi 1, 51014, Tartu, Estonia

    Keyvan Esmaeilzadeh-Salestani

Authors
  1. Mohammad Hashemabadi
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  2. Hosseinali Sasan
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  3. Mojdeh Amandadi
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  4. Keyvan Esmaeilzadeh-Salestani
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  5. Saeed Esmaeili-Mahani
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  6. Hadi Ravan
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Contributions

All authors contributed to the study’s conception and design. The design and writing of the manuscript were performed by MH and MA. Data collection was performed by MH, HS, and KE, and data analysis was performed by MH, SE, and HR.

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Correspondence to Hosseinali Sasan.

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The authors declare that they have no conflict of interest.

Ethical Approval

All experiments and procedures were approved by the Animal Research Ethics Committee of the Kerman Neuroscience Research Center, Kerman, Iran (EC/KNRC/86–31).

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Hashemabadi, M., Sasan, H., Amandadi, M. et al. CRISPR/Cas9-Mediated Disruption of ZNF543 Gene: An Approach Toward Discovering Its Relation to TRIM28 Gene in Parkinson’s Disease. Mol Biotechnol 65, 243–251 (2023). https://doi.org/10.1007/s12033-022-00494-0

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