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

被引:6
|
作者
Tokgun, Onur [1 ]
Fiorentino, Francesco Paolo [2 ,3 ]
Tokgun, Pervin Elvan [1 ]
Yokota, Jun [4 ]
Akca, Hakan [1 ]
机构
[1] Pamukkale Univ, Dept Med Biol, Sch Med, Denizli, Turkey
[2] Kitos Biotech Srls, Porto Conte Ric, I-07041 Alghero, Italy
[3] Univ Sassari, Dept Biomed Sci, Sassari, Italy
[4] Inst Invest Germans Trias & Pujol IGTP, Genom & Epigen Canc Predict Program, Campus Can Ruti, Barcelona, Spain
关键词
Lentivirus; Myc; MCS; SCLC; Dox; Inducible vector; CELL-CYCLE REGULATORS; TET-ON SYSTEM; GENE-THERAPY; VIRAL VECTORS; NERVOUS-SYSTEM; LINES;
D O I
10.1007/s12033-017-0006-y
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:200 / 206
页数:7
相关论文
共 50 条
  • [21] Construction and characterization of a novel maltose inducible expression vector in Bacillus subtilis
    Yang Ming-Ming
    Zhang Wei-Wei
    Zhang Xi-Feng
    Cen Pei-Lin
    [J]. Biotechnology Letters, 2006, 28 : 1713 - 1718
  • [22] Immortalization of erythroid precursor cells by c-myc and BCL-XL using antibiotic inducible lentiviral vector system
    Kronstein-Wiedemann, R.
    Zimmermann, S.
    Tonn, T.
    [J]. TRANSFUSION MEDICINE AND HEMOTHERAPY, 2017, 44 : 84 - 84
  • [23] Efficient control of gene expression in the hematopoietic system using a single Tet-on inducible lentiviral vector
    Barde, I
    Zanta-Boussif, MA
    Paisant, S
    Leboeuf, M
    Rameau, P
    Delenda, C
    Danos, O
    [J]. MOLECULAR THERAPY, 2006, 13 (02) : 382 - 390
  • [24] A new approach for stable lentiviral vector producer cell line development
    Tomas, H. A.
    Rodrigues, A. F.
    Formas-Oliveira, A. S.
    Alves, P. M.
    Carrondo, M. J. T.
    Coroadinha, A. S.
    [J]. HUMAN GENE THERAPY, 2015, 26 (10) : A100 - A100
  • [25] Design and Characterization of an "All-in-One" Lentiviral Vector System Combining Constitutive Anti-GD2 CAR Expression and Inducible Cytokines
    Zimmermann, Katharina
    Kuehle, Johannes
    Dragon, Anna Christina
    Galla, Melanie
    Kloth, Christina
    Rudek, Loreen Sophie
    Sandalcioglu, I. Erol
    Neyazi, Belal
    Moritz, Thomas
    Meyer, Johann
    Rossig, Claudia
    Altvater, Bianca
    Eiz-Vesper, Britta
    Morgan, Michael Alexander
    Abken, Hinrich
    Schambach, Axel
    [J]. CANCERS, 2020, 12 (02)
  • [26] Construction of a tetracycline inducible expression vector and characterization of its use in Vibrio cholerae
    Bina, X. Renee
    Wong, Eileen A.
    Bina, Thomas F.
    Bina, James E.
    [J]. PLASMID, 2014, 76 : 87 - 94
  • [27] Construction and characterization of a zinc-inducible gene expression vector in fission yeast
    Takahata, Shinya
    Asanuma, Takahiro
    Mori, Miyuki
    Murakami, Yota
    [J]. YEAST, 2021, 38 (04) : 251 - 261
  • [28] Inducible expression based on regulated recombination: a single vector strategy for stable expression in cultured cells
    Angrand, PO
    Woodroofe, CP
    Buchholz, F
    Stewart, AF
    [J]. NUCLEIC ACIDS RESEARCH, 1998, 26 (13) : 3263 - 3269
  • [29] CONSTRUCTION OF A RECOMBINANT LENTIVIRAL VECTOR CONTAINING MIR-210 AND ITS STABLE EXPRESSION IN HMEC
    Lou Yuanlei
    Deng Zhifeng
    Gao Fangliang
    Ruan Qiangfang
    Xie An
    Guo Fei
    Wang Yang
    [J]. IFPT'6: PROGRESS ON POST-GENOME TECHNOLOGIES, PROCEEDINGS, 2009, : 239 - +
  • [30] Construction of lentiviral vector carrying Rab9 gene and its expression in mouse brain
    Hao Y.
    Zhang M.
    Xu J.
    Bu B.
    Wei J.
    [J]. Frontiers of Medicine in China, 2009, 3 (2): : 141 - 147