title: 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.
abstract: 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.
authors.affiliation: Diagnostic and Molecular Medicine Healthcare Group, Box 113 Veterans Affairs Medical Center, 5901 E. 7th Street, Long Beach, CA 90822, USA. jiangang.zhang@med.va.gov
authors.forename: Jian-Gang
authors.initials: JG
authors.lastname: Zhang
authors.forename: Qinghong
authors.initials: Q
authors.lastname: Dan
authors.forename: Timothy C
authors.initials: TC
authors.lastname: Fong
authors.forename: Christopher C
authors.initials: CC
authors.lastname: Williams
authors.forename: Maria D
authors.initials: MD
authors.lastname: Avina
authors.forename: Mehrdokht
authors.initials: M
authors.lastname: Tarbiyat-Boldaji
authors.forename: Sakineh
authors.initials: S
authors.lastname: Khalaghizadeh
authors.forename: Michael
authors.initials: M
authors.lastname: Irwin
authors.forename: Amy
authors.initials: A
authors.lastname: Nguyen
authors.forename: Jing-Li
authors.initials: JL
authors.lastname: Zhuang
authors.forename: Neil
authors.initials: N
authors.lastname: Hoa
authors.forename: H Terry
authors.initials: HT
authors.lastname: Wepsic
authors.forename: Martin R
authors.initials: MR
authors.lastname: Jadus
mesh_terms: D020121:5' Untranslated Regions / Q000235:genetics*; D000818:Animals; D002417:Cattle; D002448:Cell Adhesion; D002478:Cells, Cultured; D018432:Drug Resistance, Multiple; D005822:Genetic Vectors; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D016173:Macrophage Colony-Stimulating Factor / Q000096:biosynthesis* / Q000235:genetics; D051379:Mice; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D012190:Retroviridae / Q000235:genetics*; D014161:Transduction, Genetic
chemical_list: D020121:5' Untranslated Regions; D012333:RNA, Messenger; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D016173:Macrophage Colony-Stimulating Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor