C-Terminal Binding Protein (CtBP) single isoform lentiviral stable cell lines
OHSU # 1285
These cell lines re-introduce CtBP wild type and functional mutant isoforms (both CtBP1 and 2) back into a CtBP1+2 null mouse embryonic fibroblast (MEF) line. These cells contain lentiviral-introduced protein encoding sequences that allow for stable expression of physiologic levels of CtBP isoforms under the tetracycline operon regulated promoter. This results in a cell in which either the wild type or mutant CtBP (1 or 2) is expressed in isolation, allowing study of gene specific regulatory events that are thought to be reliant upon CtBP.
These lines allow for stable, in vivo, expression of a single isoform of CtBP at near physiologic levels and allow for examination of single, specific functional point mutants in the absence of any wild type CtBP, thereby allowing study on the effects of a specific biochemical disruption on an in vivo genomic promoter and its repression/activation.
The literature lacks significant analysis of CtBP and its function at genomic promoters. CtBP is known to be important in morphogenesis and patterning and its genetic deletion produces either serious and deleterious post-natal complications or death in mice and Drosophila. CtBP has not been well studied in cell systems at genomic promoters therefore the ability to “add-back” CtBP into the null environment is a powerful technique to examine gene specific events and how those transcriptional control events are modulated by CtBP and its intrinsic biochemical functions. The wild type lines in particular will allow for study of isoforms specific effects and may be widely useful amongst investigators trying to determine the promoter specific contributions of one or both isoforms to a gene regulation event.
Dr. Madison graduated with a MD and PhD (Biochemistry and Molecular Biology) from the University of Connecticut in 1998. He completed a residency in Internal Medicine at OHSU in 2000 and an Endocrinology fellowship at OHSU in 2003. Dr. Madison is board certified in Internal Medicine and Endocrinology.
Dr. Lundblad graduated with a MD and PhD from Columbia University in 1990 and 1993 respectively. He completed a residency in Internal Medicine at OHSU in 1992 and an Endocrinology fellowship at OHSU in 1993.
Cell lines (17 in total) have been created with wild type or mutant CtBP1 (13 lines) or CtBP2 (4 lines). All have been tested by Western blot to express the correct protein. The cell lines are ready to use as frozen cell stocks. The plasmids used to make the cell lines and the viruses synthesized from the plasmid constructs used to make the cell lines are also available.
The cell lines, plasmids and viruses are available for non-exclusive licensing.
For more information, contact:
Technology Development Manager