Max-delbruck-centrum Fur Molekulare Medizin in Der Helmholtz-gemeinschaft

Max-delbrück-centrum Für Molekulare Medizin in Der Helmholtz-gemeinshcaft

Deutsches Rheuma-forschungszentrum Berlin

Mario Bunse

Wolfgang Uckert

Zsuzsanna Izsvák

Elisa Kieback

Armin Rehm

Julian Clauss

Stephen Anderton

Ulrik Stervbo

Vicky Lampropoulou

Anthea Wirges

Simon Fillatreau

The present invention relates to the field of genetic engineering, in particular, to a transposon-based transfection kit suitable for transfection of primary cells, such as T cells, comprising mRNA encoding a transposase, or reagents for generating mRNA encoding said transposase, as well as minicircle DNA comprising the transposon. The invention also relates to a nucleic acid, preferably, a DNA minicircle, comprising a transposon, wherein the transposon encodes a protein and at least one miRNA, wherein the sequences encoding the miRNA are located in an intron and expression of the protein and the miRNA is regulated by the same promoter. The invention also provides a population of cells obtainable with the method of the invention. Methods of transfection are also provided, as well as medical use, e.g. in immunotherapy, in particular, in adoptive T cell therapy or T cell receptor (TCR) or chimeric antigen receptor (CAR) gene therapy.

Transposon-based transfection system for primary cells

Growing community of inventors

Berlin, Germany

Mario Bunse