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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., Gogvadze, E., Kholodenko, R., Grzela, D.P., Mityaev, M., Vinogradova, T., Kopantzev, E., Malakhova, G., Suntsova, M., Sokov, D., 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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