title: A flexible approach to studying post-transcriptional gene regulation in stably transfected mammalian cells.
abstract: The study of post-transcriptional regulation is constrained by the technical limitations associated with both transient and stable transfection of chimeric reporter plasmids examining the activity of 3'-UTR cis-acting elements. We report the adaptation of a commercially available system that enables consistent stable integration of chimeric reporter cDNA into a single genomic site in which transcription is induced by tetracycline. Using this system, we demonstrate the tight control afforded by this system and its suitability in mapping the regulatory function of defined cis-acting elements in the human TNF 3'-UTR, as well as the distinct effects of serum starvation on transiently transfected and stably integrated chimeric reporter genes.
authors.affiliation: Veterans Administration Research Service, Veterans Affairs Medical Center, Mailstop 151, 215 North Main Street, White River Junction, VT 05009-0001, USA. ralph.c.nichols@dartmouth.edu
authors.forename: Ralph C
authors.initials: RC
authors.lastname: Nichols
authors.forename: John
authors.initials: J
authors.lastname: Botson
authors.forename: Xiao Wei
authors.initials: XW
authors.lastname: Wang
authors.forename: B JoNell
authors.initials: BJ
authors.lastname: Hamilton
authors.forename: Jane E
authors.initials: JE
authors.lastname: Collins
authors.forename: Victoria
authors.initials: V
authors.lastname: Uribe
authors.forename: Seth A
authors.initials: SA
authors.lastname: Brooks
authors.forename: Moe
authors.initials: M
authors.lastname: Zan
authors.forename: William F C
authors.initials: WF
authors.lastname: Rigby
mesh_terms: D020413:3' Untranslated Regions; D001483:Base Sequence; D003627:Data Interpretation, Statistical; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D008957:Models, Genetic; D008967:Molecular Biology / Q000379:methods*; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D050436:Regulatory Elements, Transcriptional; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000235:genetics
chemical_list: D020413:3' Untranslated Regions; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D049452:Green Fluorescent Proteins