mRNA Transfection of T-Lymphocytes by Electroporation
- 375 Downloads
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 wordsElectroporation mRNA transfection CD4+-T cells isolation Gene editing CCR5-knockout Gene therapy
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.
- 5.Tsong TY (1990) On electroporation of cell membranes and some. Relat Phenom 299:271–295Google Scholar
- 10.Aksoy P, Aksoy BA, Czech E, Hammerbacher J (2018) Electroporation characteristics of human primary T cells bioRxiv 466243. https://doi.org/10.1101/466243
- 17.Cesana D, Volpin M, Serina Secanechia YN, Montini E (2017) Safety and efficacy of retroviral and lentiviral vectors for gene therapy. In: Brunetti-Pierri N (ed) Safety and efficacy of gene-based therapeutics for inherited disorders. Springer International Publishing, Cham, pp 9–35CrossRefGoogle Scholar
- 18.Porteus M (2016) Genome editing: a new approach to human therapeutics. Annu Rev Pharmacol Toxicol 56:163–190. https://doi.org/10.1146/annurev-pharmtox-010814-124454CrossRefPubMedGoogle Scholar