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Abstract <p>Advances in recombinant technologies developed in the middle of the last century have made it possible to effectively combat diseases such as insulin-dependent diabetes mellitus. Despite the passage of time, however, insulin injections remain the primary therapeutic modality. The advent of contemporary cell technologies and genome engineering has paved the way for the development of less invasive methodologies predicated on the long-term transplantation of insulin-producing cells. In this study, we investigated the possibility of creating a line of induced pluripotent stem cells into which a genetic construct was introduced using the CRISPR/Cas9 genome editing method. The genetic construct under investigation contained the human immune checkpoint gene programmed death-ligand 1 PD-L1 under the control of the insulin promoter. The safe harbor adeno-associated virus integration site 1, located on chromosome 19, was selected as the transgene integration site. The studies confirmed the possibility of creating such a line and expressing the transgene.</p>

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Development of a System for the Safe Insertion of Transgenes into the Genomes of Model Cell Lines and Pluripotent Stem Cells

  • 期刊论文

  • 发表日期：

    2026年4月15日

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  • A. V. Selezneva,
  • A. O. Sosnovtseva,
  • E. S. Starodubova,
  • D. O. Antonov,
  • V. I. Barkovskaya,
  • S. L. Kiselev

摘要

Abstract

Advances in recombinant technologies developed in the middle of the last century have made it possible to effectively combat diseases such as insulin-dependent diabetes mellitus. Despite the passage of time, however, insulin injections remain the primary therapeutic modality. The advent of contemporary cell technologies and genome engineering has paved the way for the development of less invasive methodologies predicated on the long-term transplantation of insulin-producing cells. In this study, we investigated the possibility of creating a line of induced pluripotent stem cells into which a genetic construct was introduced using the CRISPR/Cas9 genome editing method. The genetic construct under investigation contained the human immune checkpoint gene programmed death-ligand 1 PD-L1 under the control of the insulin promoter. The safe harbor adeno-associated virus integration site 1, located on chromosome 19, was selected as the transgene integration site. The studies confirmed the possibility of creating such a line and expressing the transgene.