Cloning of human SOX2 in lentiviral vector and transduction of HEK293T cell line

Recently the regenerative stem cell-based therapy has held great promise in treating severe degenerative diseases. SOX2 gene is highly conserved among species and plays an important role in maintenance of self renewal capacity in stem cells. SOX2 concomitant with OCT4، cMYC، and KLF4 is recruited to reprogram somatic cells towards stem cells. Today SOX2 is frequently being used in induced pluripotent stem cells generation and desired cell based therapies. The aim of this study was to clone SOX2 gene in lentiviral vector and evaluate HEK293T transduction. In the present experimental study، the coding sequence of human SOX2 gene was synthesized and received in pUC57 cloning vector. The synthesized cDNA was sub-cloned into pLEX- MCS Lentiviral vector. Lentiviral particles were produced in HEK293T cells and subsequently the HEK293T were transducted and infected cells were selected by their resistance to puromycin in the culture. Expression of SOX2 was evaluated in infected HEK293 cells by using Real-Time PCR. Constructs expressing SOX2 gene were produced and confirmed. HEK293T was successfully transducted by lentiviral particles and after 24 hours more than 90% of HEK293T represented the expression of GFP. Real-Time PCR confirmed SOX2 expression in infected HEK293 cells. HEK293T wase transducted and expressed GFP and SOX2 successfully.