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Abstract <p><b>Objective:</b> Scientists need cell models from human tissues to develop methods of gene therapy and genome editing for monogenic diseases. It is preferable to use minimally invasive methods to obtain samples; these tissues can be applied for further screening to select the most effective approach to restore the synthesis of the target protein. <b>Methods:</b> We used the CRISPR/Cas9-SAM transcriptional activation system, which ensures expression of the <i>DYSF</i> gene in HEK293Т cells, as well as in fibroblasts from patients with dysferlinopathy (c.2779delG (Ala927LeufsX21)). <b>Results and Discussion:</b> After targeted activation of <i>DYSF</i>, it was possible to detect the main gene products: mRNA and protein (HEK293Т_ТА) and mRNA (fibroblasts). <b>Conclusions:</b> Transcriptionally activated dysferlin-deficient fibroblasts and HEK293 cells can be used to evaluate the in vitro efficacy of gene therapy for dysferlinopathies.</p>

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Development of an In Vitro Model of Dysferlinopathy via CRISPR/Cas-Mediated Transcriptional Activation of the DYSF Gene

  • 期刊论文

  • 发表日期：

    2025年4月3日

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  • I. A. Yakovlev,
  • Y. S. Slesarenko,
  • I. G. Starostina,
  • A. A. Shaimardanova,
  • V. V. Solovyova,
  • P. A. Bobrovsky,
  • E. N. Grafskaia,
  • L. D. Belikova,
  • S. N. Bardakov,
  • A. A. Rizvanov,
  • A. A. Isaev,
  • R. V. Deev

摘要

Abstract

Objective: Scientists need cell models from human tissues to develop methods of gene therapy and genome editing for monogenic diseases. It is preferable to use minimally invasive methods to obtain samples; these tissues can be applied for further screening to select the most effective approach to restore the synthesis of the target protein. Methods: We used the CRISPR/Cas9-SAM transcriptional activation system, which ensures expression of the DYSF gene in HEK293Т cells, as well as in fibroblasts from patients with dysferlinopathy (c.2779delG (Ala927LeufsX21)). Results and Discussion: After targeted activation of DYSF, it was possible to detect the main gene products: mRNA and protein (HEK293Т_ТА) and mRNA (fibroblasts). Conclusions: Transcriptionally activated dysferlin-deficient fibroblasts and HEK293 cells can be used to evaluate the in vitro efficacy of gene therapy for dysferlinopathies.