Biopolym. Cell. 2017; 33(5):356-366.
http://dx.doi.org/10.7124/bc.00095F
Молекулярна та клітинна біотехнології
Створення стабільних ліній клітин HEK-293 з порушеною експресією ізоформ кінази рибосомного білка S6 (S6K1) за допомогою системи редагування генома CRISPR/Cas9
1Заєць І. В., 1Сівченко А. С., 1Хоруженко А. І., 1Філоненко В. В.
  1. Інститут молекулярної біології і генетики НАН України
    вул. Академіка Заболотного, 150, Київ, Україна, 03680

Abstract

Мета. Створити клітини HEK293 з порушеною експресією S6K1 ізоформ: p85, p70 і Р60. Методи. CRISPR/cas9 система редагування генома, Вестерн-блот, імунофлуоресцентний аналіз, ПЛР-аналіз, MTT тест, метод «раневої поверхні». Результати. Отримано декілька клонів клітин НЕК-293 з повною втратою експресії білка p85/p70/p60-S6K1. Було оцінено ефект нокауту p85/p70/p60-S6K1 на Akt/mTORC1/S6K1 сигналінг та клітинні проліферацію і міграцію. Висновки. Створені клітинні лінії можуть бути використані для вивчення ролі, яку відіграє S6K1 в фізіології клітини, що дозволяє отримати більш детальне уявлення про клітинні функції ізоформ S6K1. Клітини / / HEK-293 виявляють знижений рівень фосфорилювання Akt по Сер473 і подальше пригнічення швидкості клітинного росту разом з інгібуванням клітинної рухливості.
Повний текст: (PDF, англійською)

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