Cloning of hsa-miR-26a and Construction of Its Lentiviral Expression Vector

Objective To construct a lentiviral expression vector for hsa-miR-26a. Methods The pre-mir-26a-1 amplified by PCR was inserted into plVTHM. The recombinant plasmid plVTHM-miR-26a was confirmed by restriction endonuclease analysis and DNA sequencing. 293FT cells were cotransfected with lentiviral vector plVTHM-miR-26a, psPAX2 and pMD2.G. All virus stocks were produced by calcium phosphate-mediated transfection. Virus titer was measured according to the expression level of GFP. Results Restriction enzyme digestion and DNA sequencing demonstrated that the lentivirus vector plVTHM-miR-26a was constructed successfully. Virus titer was 5×105 TU/ml. Conclusion The successful construction of lentivirus vector plVTHM-miR-26a provides the basis for further study of molecular functions of miR-26a.