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Easy and robust electrotransfection protocol for efficient ectopic gene expression and genome editing in human B cellsстатья Исследовательская статья
Статья опубликована в высокорейтинговом журнале
Статья опубликована в журнале из списка Web of Science и/или Scopus- Авторы: Canoy Reynand Jay, André Franck, Shmakova Anna, Wiels Joëlle, Lipinski Marc, Vassetzky Yegor, Germini Diego
- Журнал: Gene Therapy
- Год издания: 2020
- Издательство: Nature Publishing Group
- Местоположение издательства: United Kingdom
- DOI: 10.1038/s41434-020-00194-x
- Аннотация: B-cell lines and primary PBMCs are notoriously hard to transfect, thus making genome editing, ectopic gene expression, or gene silencing experiments particularly tedious. Here we propose a novel efficient and reproducible protocol for electrotransfection of lymphoblastoid, B-cell lymphoma, leukemia cell lines, and B cells from PBMCs. The proposed protocol requires neither costly equipment nor expensive reagents; it can be used with small or large plasmids. Transfection and viability rates of about 79% and 58%, respectively, have been routinely achieved by optimizing the salt concentration in the electrotransfection medium and the amount of plasmid used. A validation of the protocol was obtained via the generation of a TP53−/− RPMI8866 lymphoblastoid cell line which should prove useful in future hematological and blood cancer studies.
- Добавил в систему: Шмакова Анна Андреевна