Easy and robust electrotransfection protocol for efficient ectopic gene expression and genome editing in human B cells

Gene Ther. 2023 Feb;30(1-2):167-171. doi: 10.1038/s41434-020-00194-x. Epub 2020 Sep 30.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Ectopic Gene Expression*
  • Gene Editing* / methods
  • Humans
  • Plasmids
  • Transfection