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

BMC Biotechnology

Research Article

Zoltán Ivics¹  Dawid P Grzela¹  Dmitry Sokov²  Eugene Kopantzev³  Elena Gogvadze³  Maxim Mityaev³  Anton Buzdin³  Tatyana Vinogradova³  Maria Suntsova³  Roman Kholodenko³  Galina Malakhova³  Denis Kuzmin³ 

[1] Max Delbrück Center for Molecular Medicine, Berlin, Germany;P.A. Herzen Moscow Oncological Research Institute, Moscow, Russia;Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow, Russia;

关键词: Suicide Gene;    Testicular Germ Cell;    Sleep Beauty;    Herpes Simplex Virus Thymidine Kinase;    Suicide Gene Therapy;   

DOI  :  10.1186/1472-6750-10-58

 received in 2010-03-29, accepted in 2010-08-17,  发布年份 2010

来源: Springer

【 摘 要 】

BackgroundTissue 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.ResultsAmong 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.ConclusionsWe 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.

【 授权许可】

Unknown   
© Kuzmin et al; licensee BioMed Central Ltd. 2010. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.