Production of engineered t-cells by sleeping beauty transposon coupled with methotrexate selection

摘要

Aspects of the invention described herein include methods of treating, inhibiting, ameliorating and/or eliminating a virus or cancer cells in a subject utilizing genetically engineered human T-cells having receptors for a molecule presented by the virus or the cancer cells, wherein the genetically engineered T cells are isolated utilizing a two-stage MTX selection that employs increasing concentrations of MTX. Aspects of the invention further include a gene delivery polynucleotide, wherein the gene delivery polynucleotide is a minicircle. The minicircle may comprise a transposon. The polynucleotide may further comprise a mutant of dihydrofolate reductase. Aspects of the invention further include methods of using the gene delivery polynucleotides disclosed herein to create transformed T-cells that may be screened with methotrexate.