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Simultaneous knockdown of the expression of two genes using multiple shRNAs and subsequent knock-in of their expressionстатья
Статья опубликована в высокорейтинговом журнале
Информация о цитировании статьи получена из
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Scopus
Статья опубликована в журнале из списка Web of Science и/или Scopus
Дата последнего поиска статьи во внешних источниках: 10 августа 2018 г.
- Авторы: Xue-Ming Xu, Yoo Min-Hyuk, Carlson Bradley A., Gladyshev Vadim N., Hatfield Dolph L.
- Журнал: Nature Protocols
- Том: 4
- Номер: 9
- Год издания: 2009
- Издательство: Nature Publishing Group
- Местоположение издательства: United Kingdom
- Первая страница: 1338
- Последняя страница: 1348
- DOI: 10.1038/nprot.2009.145
- Аннотация: Small hairpin RNA (shRNA) is a powerful tool for inhibiting gene expression. One limitation has been that this technique has been used primarily to target a single gene. This protocol expands upon previous methods by describing a knockdown vector that facilitates cloning of multiple shRNAs; this allows targeted knockdown of more than one gene or of a single gene that may otherwise be difficult to knockdown using a single shRNA. The targeted gene(s) can be readily re-expressed by transfecting knockdown cells with a knock-in vector, containing an shRNA-refractive cDNA that will express the protein-of-interest even in the presence of shRNAs. The constructed knockdown and knock-in vectors can be easily used concurrently to assess possible interrelationships between genes, the effects of gene loss on cell function and/or their restoration by replacing targeted genes one at a time. The entire knockdown or knock-in procedure can be completed in ∼3–4 months.
- Добавил в систему: Гладышев Вадим Николаевич