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Novel strong tissue specific promoter for gene expression in human germ cells

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Item Type:Article
Title:Novel strong tissue specific promoter for gene expression in human germ cells
Creators Name:Kuzmin, D. and Gogvadze, E. and Kholodenko, R. and Grzela, D.P. and Mityaev, M. and Vinogradova, T. and Kopantzev, E. and Malakhova, G. and Suntsova, M. and Sokov, D. and Ivics, Z. and Buzdin, A.
Abstract:BACKGROUND: Tissue specific promoters may be utilized for a variety of applications, including programmed gene expression in cell types, tissues and organs of interest, for developing different cell culture models or for use in gene therapy. We report a novel, tissue-specific promoter that was identified and engineered from the native upstream regulatory region of the human gene NDUFV1 containing an endogenous retroviral sequence. RESULTS: Among seven established human cell lines and five primary cultures, this modified NDUFV1 upstream sequence (mNUS) was active only in human undifferentiated germ-derived cells (lines Tera-1 and EP2102), where it demonstrated high promoter activity (~twice greater than that of the SV40 early promoter, and comparable to the routinely used cytomegaloviral promoter). To investigate the potential applicability of the mNUS promoter for biotechnological needs, a construct carrying a recombinant cytosine deaminase (RCD) suicide gene under the control of mNUS was tested in cell lines of different tissue origin. High cytotoxic effect of RCD with a cell-death rate ~60% was observed only in germ-derived cells (Tera-1), whereas no effect was seen in a somatic, kidney-derived control cell line (HEK293). In further experiments, we tested mNUS-driven expression of a hyperactive Sleeping Beauty transposase (SB100X). The mNUS-SB100X construct mediated stable transgene insertions exclusively in germ-derived cells, thereby providing further evidence of tissue-specificity of the mNUS promoter. CONCLUSIONS: We conclude that mNUS may be used as an efficient promoter for tissue-specific gene expression in human germ-derived cells in many applications. Our data also suggest that the 91 bp-long sequence located exactly upstream NDUFV1 transcriptional start site plays a crucial role in the activity of this gene promoter in vitro in the majority of tested cell types (10/12), and an important role - in the rest two cell lines.
Keywords:Tumor Cell Line, Molecular Cloning, DNA Transposable Elements, Gene Expression, Gene Transfer Techniques, Transgenic, Suicide Genes, Genetic Engineering, Germ Cells, NADH Dehydrogenase, Organ Specificity, Genetic Promoter Regions, Transgenes
Source:BMC Biotechnology
Publisher:BioMed Central
Page Range:58
Date:17 August 2010
Official Publication:https://doi.org/10.1186/1472-6750-10-58
PubMed:View item in PubMed

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