Recombinant pCDH-miR-451 Lentivirus Production for the Differentiation of Stem Cells into Erythroid Cells

BACKGROUND Lentiviral expression vectors are the most effective vehicles for the delivery and expression of a gene of interest to almost any mammalian cell including non-dividing cells and model organisms. Recent studies have demonstrated that by packaging the lentivector construct into viral particles, we can obtain highly efficient transduction of expression constructs. METHODS The pCDH-451 plasmid was produced by ligation of 250 bp fragments encompassing pri-miR-451 sequences, into the XbaI /BamHI restriction sites of pCDH-CMV-MCS-EF1-copGFP vector. PAX2 plasmid (containing gag and pol genes) and pMD2 plasmid (containing vsv gene) were co-transfected with pCDH-451 plasmid. The number of viruses in the functional copy was detected using GFP protein and fluorescent microscope forty-eight hours later. RESULTS Approximately 95 % of cells in pCDH-451 groups with green fluorescent were distinguished 48 and 72 hours after infection. GFP Overexpression of lentiviruses in HEK-293T cells was 81.48% in cells transfected with 16 μl lentiviral vector expressing pCDHmiR-451. CONCLUSTION Our results demonstrate that the titer of pCDH-miR-451 lentiviruses was excellent and if stem cells were transduced with these viruses, mature miR-451 expression level are increased, and the stem cells differentiated into erythroid lineage .

Stem Cells, pCDH-451, Lentiviruses, Erythroid Differentiation