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<p><b>Abstract</b>—<b>Objective.</b> New advances in genomic editing field have led to the creation of DNA base editors. Such a tool allows to make precise changes to the genome without double-stranded DNA breaks, which reduces its mutagenicity. As a result, DNA base editors are becoming a popular tool for gene therapy and transgenesis of cell lines and laboratory animals. Certainly, there is a need not only to evaluate the effectiveness of editing, but also to create new forms of DNA base editors through evolution. The purpose of our work is to create system to evaluate the effectiveness of DNA base editors and their evolution to obtain new forms. <b>Materials and methods.</b> Classical methods of genetic engineering were used in the work. The proportion of cells with the target effect was assessed by flow cytometry. <b>Results.</b> It is shown that the created systems simulate transgene expression depending on the state of its start-codons. It was revealed that editing of the first start-codon plays a key role in the initiation of translation or its inhibition. <b>Conclusions.</b> Based on the developed systems, it is possible to evaluate the effectiveness of editing the first start-codon of the HIV-1 <i>gag</i> gene and the evolution of DNA base editors to obtain new forms.</p>

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Creation of System to Evaluate the Effectiveness of DNA Base Editors and Its Evolution

  • 期刊论文

  • 发表日期：

    2025年12月1日

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  • T. I. Aliev,
  • E. Yu. Prudnikova,
  • A. R. Imatdinov,
  • I. R. Imatdinov

摘要

Abstract—Objective. New advances in genomic editing field have led to the creation of DNA base editors. Such a tool allows to make precise changes to the genome without double-stranded DNA breaks, which reduces its mutagenicity. As a result, DNA base editors are becoming a popular tool for gene therapy and transgenesis of cell lines and laboratory animals. Certainly, there is a need not only to evaluate the effectiveness of editing, but also to create new forms of DNA base editors through evolution. The purpose of our work is to create system to evaluate the effectiveness of DNA base editors and their evolution to obtain new forms. Materials and methods. Classical methods of genetic engineering were used in the work. The proportion of cells with the target effect was assessed by flow cytometry. Results. It is shown that the created systems simulate transgene expression depending on the state of its start-codons. It was revealed that editing of the first start-codon plays a key role in the initiation of translation or its inhibition. Conclusions. Based on the developed systems, it is possible to evaluate the effectiveness of editing the first start-codon of the HIV-1 gag gene and the evolution of DNA base editors to obtain new forms.