Эффективный перенос трансгена в лейкозные клетки человека с помощью лентивирусного вектора второго поколения

Abstract

We have found that three-vectors system based on envelope plasmid pMD2.G and packaging plasmid pCMV_dR8.91 as well as lentivirus transfer vector pHR-SINcPPt-SIEW permits to obtain recombinant pseudotyped lentiviruses with titer at least 105 transducing unites per 1 ml of native supernatant from lentivirus-producing 293T cells. According to both flow cytometry and fluorescence microscopy data, at such functional lentivirus titer the efficiency of transfer of the egfp reporter gene (as a part of the lentiviral vector pHR-SINcPPt-SIEW) into Kasumi-1 acute myeloid leukemia cells and SEM acute B-lymphoblastic leukemia cells is higher than 80 % and higher than 90 %, respectively. After 10-fold concentration of lentivirus-containing supernatant the efficiency of transduction of leukemia cells was increased up to approximately 100 %. This data indicate that the pHR-SINcPPt-SIEW self-inactivating lentivirus transfer vector can be considered as a powerful tool for gene modification of leukemia cells. = Лентивирусный вектор второго поколения pHR-SINcPPt-SIEW позволяет модифицировать до 100 % клеток перевиваемых лейкозных клеточных линий человека и может рассматриваться как эффективный инструмент в генетической модификации лейкозных клеток.