Abstract
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.
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Acknowledgements
This research was supported by grants from the GEFLUC, LNCC, the Presidium of RAS and the State program of fundamental scientific research of IDB RAS. We gratefully acknowledge the technical support of the imaging and cytometry platform of the Gustave Roussy Institute.
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Canoy, R.J., André, F., Shmakova, A. et al. Easy and robust electrotransfection protocol for efficient ectopic gene expression and genome editing in human B cells. Gene Ther 30, 167–171 (2023). https://doi.org/10.1038/s41434-020-00194-x
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DOI: https://doi.org/10.1038/s41434-020-00194-x
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