Macrophage colony-stimulating factor expression in retrovirally transduced cells is dependent upon both the adherence status of the target cells and its 5′ flanking untranslated region

• Published in: Biochemical and biophysical research communications 2005-05, Vol.330 (4), p.1275-1284
• Main Authors: Zhang, Jian-Gang, Dan, Qinghong, Fong, Timothy C., Williams, Christopher C., Avina, Maria D., Tarbiyat-Boldaji, Mehrdokht, Khalaghizadeh, Sakineh, Irwin, Michael, Nguyen, Amy, Zhuang, Jing-Li, Hoa, Neil, Wepsic, H. Terry, Jadus, Martin R.
• Format: Article
• Language: English
• Subjects: 5' Untranslated Regions - genetics; Animals; Cattle; Cell Adhesion; Cells, Cultured; Drug Resistance, Multiple; Genetic Vectors; Granulocyte-Macrophage Colony-Stimulating Factor - genetics; Green Fluorescent Proteins - biosynthesis; Green Fluorescent Proteins - genetics; Humans; M-CSF promoter; Macrophage colony-stimulating factor; Macrophage Colony-Stimulating Factor - biosynthesis; Macrophage Colony-Stimulating Factor - genetics; Mice; Rats; Real-time PCR; Retroviral transduction; Retroviridae - genetics; RNA, Messenger - biosynthesis; Transduction, Genetic
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2005.03.113

Numerous cell types retrovirally transduced with macrophage colony-stimulating factor (M-CSF) using LXSN-based vectors showed a variable expression of the transgene. Expression of M-CSF correlated with the cells’ adherent status. Transduced adherent cells produced the M-CSF, whereas the non-adherent cells synthesized little M-CSF. Studies showed that the 5′-UTR of the M-CSF gene regulated transgenic M-CSF gene expression. Ligation of this 5′-UTR to the enhanced green fluorescent protein gene (EGFP) caused the expression of EGFP to show the same dichotomy as previously seen with the M-CSF. Transgenic M-CSF was expressed within non-adherent cells when the 5′-UTR was removed from the LXSN vector. Quantitative real-time polymerase chain reaction analysis confirmed that lesser production of M-CSF mRNA occurred within the non-adherent cells than in the adherent cells. This difference was eliminated when the 5′-UTR was removed from the retroviral vector. Our work suggests that this 5′-UTR of the M-CSF gene could be an important way to get transgenic expression within adherent cells, but not in non-adherent cells.