Методы маркирования клеток для изучения судьбы клеточных поколений

В процессе развития многоклеточного организма из одной тотипотентной зиготы образуется огромное количество клеток (триллионы для человека) с разной специализацией. Во взрослом состоянии многие ткани постоянно самообновляются: старые клетки погибают, а из популяции стволовых клеток образуются новые. Изучение судьбы отдельных клеток, их происхождения и родственных связей дает ответы на многие вопросы, связанные как с нормальным развитием, так и с патогенезом. Прямые наблюдения за развивающимися эмбрионами позволили установить судьбу бластомеров асцидии Styela partita, а также происхождение всех клеток червя Caenorhabditis elegans. Исследователям повезло с объектом: в первом случае происходит естественное «маркирование » бластомеров, во втором, поскольку тело червя прозрачно, можно следить за каждой клеткой нематоды. В большинстве же случаев определение клеточных линий и идентификация субпопуляций стволовых клеток представляют большую трудность для исследователей. Поэтому для отслеживания судьбы отдельных клеток стали разрабатываться методы маркирования, основанные на внесении в клетки специфических меток, которые наследуются в ходе делений. Поскольку все потомки исходной клетки несут одинаковые метки, их можно легко отличить от потомков других клеток. В обзоре обсуждаются методы маркирования клеток с помощью красителей и генетических конструкций, обеспечивающих синтез белков-репортеров, по наличию которых можно установить родство клеток. Особое внимание уделено методам, основанным на внесении наследуемой метки в геном клеток (генетическое маркирование), в том числе вирусному маркированию и клеточному баркодированию. Один из разделов обзора посвящен маркированию клеток с помощью системы CRISPR/Cas – популярного инструмента генной инженерии.

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