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T-Helper Cells pp 217-226 | Cite as

mRNA Transfection of T-Lymphocytes by Electroporation

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Part of the Methods in Molecular Biology book series (MIMB, volume 2285)

Abstract

Electroporation enables the transfection of different cell types including microbial, plant, and animal cells with charged molecules, such as nuclear acids or proteins. During electroporation, an electrical field is applied to the cells leading to a transient permeabilization of the cell membrane allowing exogenous molecules to enter the cells. Here we report the electroporation of human primary CD4+ -T cells with in-vitro transcribed mRNA to facilitate gene editing (knockout) of the CC-chemokine receptor 5 (CCR5), the coreceptor of the human immunodeficiency virus 1 (HIV1) predominantly used during primary infection. Using such strategy of transient expression of a CCR5-specific Transcription-activator-like-effector nuclease (TALEN), we aim to protect helper T cells from de novo HIV infection.

Key words

Electroporation mRNA transfection CD4+-T cells isolation Gene editing CCR5-knockout Gene therapy 

Notes

Acknowledgments

Our work has been supported by the German Center for Infection Research (DZIF) within TTU HIV (04.803). We wish to thank Tanja Sonntag, the UKE Cytometry & Cell Sorting Facility and the Institute for Transfusion Medicine of the UKE for expert technical assistance and support.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2021

Authors and Affiliations

  1. 1.Research Department of Cell and Gene Therapy, Department of Stem Cell TransplantationUniversity Medical Center Hamburg-EppendorfHamburgGermany
  2. 2.German Center for Infection Research (DZIF)HamburgGermany

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