Методы геномного редактирования дрожжей: история и современное состояние

Дрожжи являются модельным эукариотическим организмом, на котором отрабатываются многие предположения о работе генома, а также методы его редактирования. Наиболее часто в исследовательских работах используют Saccharomyces cerevisiae, которые очень хорошо приспособлены физиологически к культивированию в условиях биореактора и признаны абсолютно безопасными. В последнее десятилетие методы генетической инженерии дрожжей претерпели значительные изменения. Появились новые инструменты, которые пришли из смежных направлений и позволили значительно ускорить процесс получения новых штаммов. Прежде всего это белки для направленного внесения изменений в последовательность ДНК. Длительное время методы редактирования генома дрожжей базировались на использовании их собственной системы гомологичной рекомбинации. Она удобна и удовлетворяла потребности исследователей на протяжении нескольких десятилетий, до того времени, когда на первый план стали выходить высокопроизводительные методы. Во втором десятилетии XXI века произошло бурное развитие высокопроизводительных подходов, в первую очередь методов анализа в биологии: геномики, транскриптомики, протеомики, метаболомики, интерактомики и др. Сформировалась биоинформационная база, которая позволила быстро обрабатывать растущий поток информации и моделировать клеточные процессы. В результате скорость анализа и предсказания мишеней для редактирования генома стала превышать скорость их получения, что, естественно, обусловило поиск новых методов генетической инженерии. Особенно сильно этот процесс затронул работы по модификации свойств микроорганизмов. Современные задачи стали требовать не единичных модификаций, а десятков и сотен, а иногда тысяч модификаций. В результате исследователи, занимающиеся дрожжами, стали вовлекать в работу новые инструменты геномного редактирования, которые ранее развивались для изучения более сложных объектов, таких как животные, растения, клеточные линии и др. Современные методы геномной инженерии дрожжей позволяют вносить несколько модификаций в геном за один шаг. В данном обзоре рассматривается вопрос применения и перспектив дальнейшего развития методов направленного редактирования генома в инженерии дрожжей.

дрожжи, Saccharomyces cerevisiae, генетическая инженерия, ZFN, цинковые пальцы, TALENS, CRISPR/Cas, Аргонавт

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