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<article article-type="research-article" dtd-version="1.3" xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xml:lang="ru"><front><journal-meta><journal-id journal-id-type="publisher-id">vavilov</journal-id><journal-title-group><journal-title xml:lang="ru">Вавиловский журнал генетики и селекции</journal-title><trans-title-group xml:lang="en"><trans-title>Vavilov Journal of Genetics and Breeding</trans-title></trans-title-group></journal-title-group><issn pub-type="epub">2500-3259</issn><publisher><publisher-name>Institute of Cytology and Genetics of Siberian Branch of the RAS</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="doi">10.18699/vjgb-26-04</article-id><article-id custom-type="elpub" pub-id-type="custom">vavilov-4982</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group><subj-group subj-group-type="section-heading" xml:lang="ru"><subject>МОЛЕКУЛЯРНАЯ И КЛЕТОЧНАЯ БИОЛОГИЯ</subject></subj-group><subj-group subj-group-type="section-heading" xml:lang="en"><subject>MOLECULAR AND CELL BIOLOGY</subject></subj-group></article-categories><title-group><article-title>Редактирование оснований в гене AUTS2 и высокопроизводительное NGS-генотипирование клонов: стратегия создания клеточной модели</article-title><trans-title-group xml:lang="en"><trans-title>Base editing in the AUTS2 gene and high-throughput NGS genotyping of clones: a strategy for generating a cellular model</trans-title></trans-title-group></title-group><contrib-group><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0003-0305-0612</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Ян</surname><given-names>A. П.</given-names></name><name name-style="western" xml:lang="en"><surname>Yan</surname><given-names>A. P.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Новосибирск; федеральная территория «Сириус», Краснодарский край</p></bio><bio xml:lang="en"><p>Novosibirsk; Sirius Federal Territory, Krasnodar region</p></bio><email xlink:type="simple">a.yan@alumni.nsu.ru</email><xref ref-type="aff" rid="aff-1"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0001-9470-7178</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Сальников</surname><given-names>П. A.</given-names></name><name name-style="western" xml:lang="en"><surname>Salnikov</surname><given-names>P. A.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Новосибирск</p></bio><bio xml:lang="en"><p>Novosibirsk</p></bio><xref ref-type="aff" rid="aff-2"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Буздин</surname><given-names>А. А.</given-names></name><name name-style="western" xml:lang="en"><surname>Buzdin</surname><given-names>А. А.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Москва</p></bio><bio xml:lang="en"><p>Moscow</p></bio><xref ref-type="aff" rid="aff-3"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-8728-8574</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Ковальская</surname><given-names>В. А.</given-names></name><name name-style="western" xml:lang="en"><surname>Кovalskaia</surname><given-names>V. А.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Москва</p></bio><bio xml:lang="en"><p>Moscow</p></bio><xref ref-type="aff" rid="aff-4"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Мусатова</surname><given-names>Е. В.</given-names></name><name name-style="western" xml:lang="en"><surname>Musatova</surname><given-names>E. V.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Москва</p></bio><bio xml:lang="en"><p>Moscow</p></bio><xref ref-type="aff" rid="aff-5"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0001-9371-2178</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Орлов</surname><given-names>П. С.</given-names></name><name name-style="western" xml:lang="en"><surname>Orlov</surname><given-names>P. S.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Новосибирск</p></bio><bio xml:lang="en"><p>Novosibirsk</p></bio><xref ref-type="aff" rid="aff-6"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Рыжкова</surname><given-names>О. П.</given-names></name><name name-style="western" xml:lang="en"><surname>Ryzhkova</surname><given-names>О. P.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Москва</p></bio><bio xml:lang="en"><p>Moscow</p></bio><xref ref-type="aff" rid="aff-4"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0009-0004-1777-523X</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Субботовская</surname><given-names>А. И.</given-names></name><name name-style="western" xml:lang="en"><surname>Subbotovskaia</surname><given-names>A. I.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Новосибирск</p></bio><bio xml:lang="en"><p>Novosibirsk</p></bio><xref ref-type="aff" rid="aff-7"/></contrib><contrib contrib-type="author" corresp="yes"><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Сунцова</surname><given-names>М. В.</given-names></name><name name-style="western" xml:lang="en"><surname>Suntsova</surname><given-names>М. V.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Москва</p></bio><bio xml:lang="en"><p>Moscow</p></bio><xref ref-type="aff" rid="aff-8"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0003-1375-4808</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Христиченко</surname><given-names>А. Ю.</given-names></name><name name-style="western" xml:lang="en"><surname>Khristichenko</surname><given-names>А. U.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Москва</p></bio><bio xml:lang="en"><p>Moscow</p></bio><xref ref-type="aff" rid="aff-9"/></contrib><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-9425-9763</contrib-id><name-alternatives><name name-style="eastern" xml:lang="ru"><surname>Хабарова</surname><given-names>А. А.</given-names></name><name name-style="western" xml:lang="en"><surname>Khabarova</surname><given-names>А. А.</given-names></name></name-alternatives><bio xml:lang="ru"><p>Новосибирск</p></bio><bio xml:lang="en"><p>Novosibirsk</p></bio><email xlink:type="simple">khabarova@bionet.nsc.ru</email><xref ref-type="aff" rid="aff-10"/></contrib></contrib-group><aff-alternatives id="aff-1"><aff xml:lang="ru">Федеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наук; Новосибирский национальный исследовательский государственный университет; Научно-технологический университет «Сириус»<country>Россия</country></aff><aff xml:lang="en">Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences; Novosibirsk State University; Sirius University of Science and Technology<country>Russian Federation</country></aff></aff-alternatives><aff-alternatives id="aff-2"><aff xml:lang="ru">Федеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наук; Новосибирский национальный исследовательский государственный университет<country>Россия</country></aff><aff xml:lang="en">Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences; Novosibirsk State University<country>Russian Federation</country></aff></aff-alternatives><aff-alternatives id="aff-3"><aff xml:lang="ru">Первый Московский государственный медицинский университет им. И.М. Сеченова Министерства здравоохранения Российской Федерации; Национальный медицинский исследовательский центр эндокринологии им. академика И.И. Дедова; Институт биоорганической химии им. академиков М.М. Шемякина и Ю.А. Овчинникова Российской академии наук<country>Россия</country></aff><aff xml:lang="en">I.M. Sechenov First Moscow State Medical University of the Ministry of Healthcare of the Russian Federation; National Medical Research Center for Endocrinology named after Academician I.I. Dedov;  M.M. Shemyakin–Yu.A. Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences<country>Russian Federation</country></aff></aff-alternatives><aff-alternatives id="aff-4"><aff xml:lang="ru">Медико-генетический научный центр им. академика Н.П. Бочкова<country>Россия</country></aff><aff xml:lang="en">Research Centre for Medical Genetics<country>Russian Federation</country></aff></aff-alternatives><aff-alternatives id="aff-5"><aff xml:lang="ru">Центр генетики и репродуктивной медицины “Genetico”<country>Россия</country></aff><aff xml:lang="en">Center of Genetics and Reproductive Medicine “Genetico”<country>Russian Federation</country></aff></aff-alternatives><aff-alternatives id="aff-6"><aff xml:lang="ru">Федеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наук; Федеральный исследовательский центр фундаментальной и трансляционной медицины<country>Россия</country></aff><aff xml:lang="en">Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences; Federal Research Center of Fundamental and Translational Medicine<country>Russian Federation</country></aff></aff-alternatives><aff-alternatives id="aff-7"><aff xml:lang="ru">Федеральный исследовательский центр фундаментальной и трансляционной медицины<country>Россия</country></aff><aff xml:lang="en">Federal Research Center of Fundamental and Translational Medicine<country>Russian Federation</country></aff></aff-alternatives><aff-alternatives id="aff-8"><aff xml:lang="ru">Первый Московский государственный медицинский университет им. И.М. Сеченова Министерства здравоохранения Российской Федерации; Национальный медицинский исследовательский центр эндокринологии им. академика И.И. Дедова<country>Россия</country></aff><aff xml:lang="en">I.M. Sechenov First Moscow State Medical University of the Ministry of Healthcare of the Russian Federation; National Medical Research Center for Endocrinology named after Academician I.I. Dedov<country>Russian Federation</country></aff></aff-alternatives><aff-alternatives id="aff-9"><aff xml:lang="ru">Национальный медицинский исследовательский центр эндокринологии им. академика И.И. Дедова<country>Россия</country></aff><aff xml:lang="en">National Medical Research Center for Endocrinology named after Academician I.I. Dedov<country>Russian Federation</country></aff></aff-alternatives><aff-alternatives id="aff-10"><aff xml:lang="ru">Федеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наук<country>Россия</country></aff><aff xml:lang="en">Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences<country>Russian Federation</country></aff></aff-alternatives><pub-date pub-type="collection"><year>2026</year></pub-date><pub-date pub-type="epub"><day>05</day><month>03</month><year>2026</year></pub-date><volume>30</volume><issue>1</issue><fpage>72</fpage><lpage>84</lpage><permissions><copyright-statement>Copyright &amp;#x00A9; Ян A.П., Сальников П.A., Буздин А.А., Ковальская В.А., Мусатова Е.В., Орлов П.С., Рыжкова О.П., Субботовская А.И., Сунцова М.В., Христиченко А.Ю., Хабарова А.А., 2026</copyright-statement><copyright-year>2026</copyright-year><copyright-holder xml:lang="ru">Ян A.П., Сальников П.A., Буздин А.А., Ковальская В.А., Мусатова Е.В., Орлов П.С., Рыжкова О.П., Субботовская А.И., Сунцова М.В., Христиченко А.Ю., Хабарова А.А.</copyright-holder><copyright-holder xml:lang="en">Yan A.P., Salnikov P.A., Buzdin А.А., Кovalskaia V.А., Musatova E.V., Orlov P.S., Ryzhkova О.P., Subbotovskaia A.I., Suntsova М.V., Khristichenko А.U., Khabarova А.А.</copyright-holder><license license-type="creative-commons-attribution" xlink:href="https://creativecommons.org/licenses/by/4.0/" xlink:type="simple"><license-p>This work is licensed under a Creative Commons Attribution 4.0 License.</license-p></license></permissions><self-uri xlink:href="https://vavilov.elpub.ru/jour/article/view/4982">https://vavilov.elpub.ru/jour/article/view/4982</self-uri><abstract><p>Изучение молекулярных механизмов, лежащих в основе расстройств аутистического спектра (РАС), требует создания клеточных моделей, способных отражать цис-регуляторные эффекты и аллель-специфичную экспрессию генов. В настоящем исследовании мы представляем подход к получению индуцированных плюрипотентных стволовых клеток (ИПСК), модифицированных с использованием аденинового редактора оснований (ABE), для введения синонимичных однонуклеотидных замен в ген AUTS2 – кандидата на участие в патогенезе РАС. Эти замены позволяют маркировать аллели и отслеживать различия в экспрессии нормального и реорганизованного аллелей в цис-контексте. Мы разработали стратегию высокоэффективного генотипирования клонов с использованием секвенирования продуктов ПЦР (ампликонов) на платформе нового поколения (NGS). Анализ более 100 субклонов показал, что предложенный подход превосходит секвенирование по Сэнгеру по масштабируемости, чувствительности и экономичности. Мы отобрали клоны с целевыми гетерозиготными заменами, оценили уровень мозаицизма и провели фазирование с герминальными гетерозиготными вариантами, позволяющее убедиться в моноклональном происхождении клеточной линии или идентифицировать аллель, ассоциированный с мутацией. Полученные линии ИПСК маркируют разные аллели гена AUTS2, что открывает перспективу анализа влияния цис-регуляторных элементов на экспрессию гена в различных типах клеток. Результаты работы подчеркивают практическую значимость редакторов оснований и целевого NGS-генотипирования при создании клеточных моделей с однонуклеотидными заменами для фундаментальных и прикладных исследований.</p></abstract><trans-abstract xml:lang="en"><p>Studying the molecular mechanisms underlying autism spectrum disorders (ASD) requires cellular models capable of capturing cis-regulatory effects and allele-specific gene expression. In this study, we present an approach for generating induced pluripotent stem cells (iPSCs) modified using an adenine base editor (ABE) to introduce synonymous single-nucleotide substitutions in the AUTS2 gene – a candidate involved in ASD pathogenesis. These substitutions serve as allele-specific markers, enabling the tracking of expression differences between normal and rearranged alleles in a cis-regulatory context. We developed a high-efficiency strategy for genotyping clones using amplicon-based next-generation sequencing (NGS). Analysis of over 100 subclones demonstrated that this approach surpasses Sanger sequencing in scalability, sensitivity, and cost-effectiveness. We selected clones with targeted heterozygous substitutions, assessed mosaicism levels, and performed phasing with germline heterozygous variants to confirm monoclonal origin and identify the allele carrying the chromosomal rearrangement. The resulting iPSC lines mark distinct AUTS2 alleles, providing a foundation for analyzing the impact of cis-regulatory elements on gene expression across different cell types. Our findings highlight the practical value of base editors and targeted NGS genotyping in creating cellular models with single-nucleotide substitutions for both basic and applied research.</p></trans-abstract><kwd-group xml:lang="ru"><kwd>индуцированные плюрипотентные стволовые клетки (ИПСК)</kwd><kwd>CRISPR/Cas9</kwd><kwd>однонуклеотидные замены</kwd><kwd>аллель-специфичная оценка экспрессии</kwd><kwd>хромосомная перестройка</kwd></kwd-group><kwd-group xml:lang="en"><kwd>induced pluripotent stem cells (iPSCs)</kwd><kwd>CRISPR/Cas9</kwd><kwd>single-nucleotide substitutions</kwd><kwd>allele-specific expression analysis</kwd><kwd>chromosomal rearrangement</kwd></kwd-group><funding-group xml:lang="en"><funding-statement>This work was supported by the RSF grant 24-25-00152.</funding-statement></funding-group></article-meta></front><back><ref-list><title>References</title><ref id="cit1"><label>1</label><citation-alternatives><mixed-citation xml:lang="ru">Baux D., Van Goethem C., Ardouin O., Guignard T., Bergougnoux A., Koenig M., Roux A.-F. 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