Design of a Lentiviral Vector for the Inducible Expression of MYC: A New Strategy for Construction Approach

作者：Tokgun Onur; Fiorentino Francesco Paolo; Tokgun Pervin Elvan; Yokota Jun; Akca Hakan^*

来源：Molecular Biotechnology, 2017, 59(6): 200-206.

DOI：10.1007/s12033-017-0006-y

• Lentivirus
• Myc
• MCS
• SCLC
• Dox
• Inducible vector

Lentiviral vectors are powerful tools for gene expression studies. Here we report the construction of pTIJ, a vector for inducible gene expression. pTIJ was generated from pTRIPZ backbone, which is designed for the inducible expression of shRNA sequences, by the introducing of a multiple cloning site upstream of the Tet promoter and the removal of miR30 flanking sequences. To evaluate pTIJ as a tool for the inducible expression of genes of interest, we introduced MYC cDNA into pTIJ and infected two small cell lung cancer cell lines, H209 and H345. Induction of MYC expression by doxycycline was detectable in both cell lines by real-time PCR and western blot analysis. This study highlights the relevance of pTIJ vector to allow the inducible expression of any gene of interest. In our belief, pTIJ will be an extremely useful tool to simplify the generation of genetically engineered cell lines for the inducible expression of cDNA sequences in biological studies. Furthermore, we report the generation of a pTIJ-MYC vector for the inducible expression of the oncogene MYC.

• 出版日期2017-6