Method disclosed by the invention describe it is a kind of can be with unprecedented efficiency, flexibility, practicability and speed by the new technology in transgenosis stable integration to lymphocyte and other mammalian cells.The new method is based on the combination for using the transposase [such as sleeping beauty (Sleeping Beauty) transposase] that mRNA is encoded and the transposable element of minicircle dna coding.The new method can realize higher gene transfer rate, while lower than conventional method toxicity, and the conventional method is the combination of the transposase and the transposable element of Plasmid DNA coding that are encoded using Plasmid DNA.The application of the present invention includes but not limited to by the transgenosis stable integration to human T lymphocyte of encoding immune receptor (such as T cell receptor or synthesis Chimeric antigen receptor), and wherein immunity receptor has specificity to the molecule of tumor cells expression.Transposase mRNA and transposons minicircle dna can be by including but not limited to that the electrotransfer method of electroporation and nuclear transfection such as imports in lymphocyte.
