Development of a BAC vector for integration-independent and tight regulation of transgenes in rodents via the Tet system

Transgenic Research
Kai SchönigHermann Bujard

Abstract

The establishment of functional transgenic mouse lines is often limited by problems caused by integration site effects on the expression construct. Similarly, tetracycline (Tet) controlled transcription units most commonly used for conditional transgene expression in mice are strongly influenced by their genomic surrounding. Using bacterial artificial chromosome (BAC) technology in constitutive expression systems, it has been shown that integration site effects resulting in unwanted expression patterns can be largely eliminated. Here we describe a strategy to minimize unfavourable integration effects on conditional expression constructs based on a 75 kb genomic BAC fragment. This fragment was derived from a transgenic mouse line, termed LC-1, which carries the Tet-inducible genes luciferase and cre (Schönig et al. 2002). Animals of this mouse line have previously been shown to exhibit optimal expression properties in terms of tightness in the off state and the absolute level of induction, when mated to appropriate transactivator expressing mice. Here we report the cloning and identification of the transgenic LC-1 integration site which was subsequently inserted into a bacterial artificial chromosome. We demonstrate that this ve...Continue Reading

References

Nov 3, 2010·Addiction Biology·Rainer SpanagelAndreas Zimmer
Sep 5, 2012·BMC Biology·Kai SchönigDusan Bartsch
Jun 14, 2012·PloS One·Tillmann WeberDusan Bartsch
Apr 30, 2015·Journal of Cardiovascular Translational Research·Michael J FlisterRat Genome and Nomenclature Committee
Jun 17, 2015·PloS One·Nicole BäumerCarsten Müller-Tidow

Citations

Jun 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·Manfred Gossen, Hermann Bujard
Sep 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·H ShizuyaM Simon
Aug 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·C BonnerotJ F Nicolas
Jan 1, 1990·Annual Review of Cell Biology·C WilsonW J Gehring
Sep 11, 1995·Nucleic Acids Research·Udo BaronHermann Bujard
Jan 1, 1994·Reproduction, Fertility, and Development·A J ClarkF Yull
Jun 23, 1995·Science·Manfred GossenHermann Bujard
Jan 1, 1993·Developmental Neuroscience·J HermsK H Schlingensiepen
Oct 1, 1996·Proceedings of the National Academy of Sciences of the United States of America·A KistnerHermann Bujard
Nov 14, 1997·The Plant Journal : for Cell and Molecular Biology·M L WangH M Goodman
Oct 15, 1998·Nature Genetics·Y ZhangA F Stewart
Mar 31, 1999·Scandinavian Journal of Clinical and Laboratory Investigation. Supplementum·L B NielsenS G Young
Jan 5, 2000·Journal of Cellular Biochemistry·M Izumi, D M Gilbert
May 17, 2000·Proceedings of the National Academy of Sciences of the United States of America·D YuD L Court
Jun 22, 2000·Proceedings of the National Academy of Sciences of the United States of America·S UrlingerW Hillen
Apr 18, 2001·Transgenic Research·P Giraldo, L Montoliu
Oct 5, 2001·Nature Reviews. Genetics·M Lewandoski
Oct 23, 2001·Genesis : the Journal of Genetics and Development·Emilio CasanovaGünther Schütz
Jan 22, 2002·Nature Neuroscience·Ivan RodriguezPeter Mombaerts
Dec 6, 2002·Nucleic Acids Research·Kai SchönigHermann Bujard
Jul 23, 2003·Journal of the American Society of Nephrology : JASN·Anna Rachel GallagherRalph Witzgall
Jun 23, 2004·PLoS Biology·Mazahir T HasanWinfried Denk
Aug 19, 2004·American Journal of Physiology. Renal Physiology·Debra F HigginsVolker H Haase
Nov 19, 2004·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Heinz E KrestelPeter H Seeburg
Jan 10, 2006·Genesis : the Journal of Genetics and Development·Caroline BeardRudolf Jaenisch
Nov 14, 2006·Journal of Molecular Biology·Mathias Hampf, Manfred Gossen
Jan 5, 2007·BMC Neuroscience·Thomas LembergerGünther Schütz
Mar 14, 2007·Nature Medicine·Batya CohenMichal Neeman
Jun 21, 2007·PloS One·Peixin ZhuMazahir T Hasan
Feb 26, 2008·Nucleic Acids Research·Adam WilliamsDimitris Kioussis
May 10, 2008·PLoS Genetics·Hongkui ZengGeorge A Gaitanaris

Related Concepts

Tetracycline Antibiotics
Tetracyclines
Cre recombinase, Enterobacteria phage P1
Shuttle Vectors
Genome
Tetrabid
Family Dipodidae
Recombinant Transgenes
Etiology
Bacterial Artificial Chromosomes

Related Feeds

Artificial Chromosomes

Artificial chromosomes are genetically engineered chromosomes derived from the DNA of a species. Discover the latest research on artificial chromosomes here.