Система CRISPR/Cas9 – универсальный инструмент геномной инженерии

Система CRISPR/Cas9 была изначально описана как механизм адаптивного иммунитета архей и бактерий, однако после ряда модификаций она нашла активное применение в генной инженерии, благодаря способности вносить направленный ДНК-разрыв с помощью короткого программируемого 20-нуклеотидного района в направляющей молекуле РНК (single guide RNA, sgРНК). Обзор посвящен современным приложениям системы CRISPR/Cas9 в генной инженерии. В первой его части описан основной механизм действия CRISPR/Cas9, особо уделено внимание причинам неспецифической активности Cas9 (off-targets). Она выражается в связывании комплексом Cas9-sgРНК нецелевых геномных участков, имеющих лишь частичную гомологию с sgРНК, что может приводить к нежелательным мутациям в геноме. В обзоре обсуждаются недавние улучшения специфичности связывания Cas9 и подходов по расширению функций CRISPR/Cas9 для трансгенеза. Популярность системы CRISPR/Cas9 в основном обусловлена ее выдающимся потенциалом для генной терапии и геномной инженерии, и последние достижения в этих областях представлены в нашем обзоре. В частности, CRISPR/Cas9 была недавно использована для контроля заражения клеток ВИЧ (вирусом иммунодефицита человека) и исправления генетических нарушений, таких как мышечная дистрофия Дюшенна и пигментный ретинит, на культурах клеток и животных моделях. Программируемость CRISPR/Cas9 облегчает создание трансгенных организмов с направленными генными мутациями, встройками генов и крупными хромосомными перестройками. Система CRISPR/Cas9 оказалась особенно востребованной для пронуклеарной микроинъекции при получении трансгенных сельскохозяйственных животных в биотехнологии. Один из разделов обзора посвящен генетическим скринингам на основе CRISPR/Cas9, которые приводят к высокоэффективной идентификации новых генов и генных сетей во многих биологических процессах. Наконец, в обзоре рассматривается технология генных драйверов, основанная на СRISPR/Cas9, которая представляет собой мощный инструмент для модификации экосистем в обозримом будущем.

СRISPR/Cas9, редактирование генома, трансгенез, клеточная терапия, мутагенная цепная реакция,

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