Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (184K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Zhang, Z.
Right arrow Articles by Lutz, B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Zhang, Z.
Right arrow Articles by Lutz, B.
Related Collections
Right arrow Recombination
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2002, Vol. 30, No. 17 e90
© 2002 Oxford University Press

Cre recombinase-mediated inversion using lox66 and lox71: method to introduce conditional point mutations into the CREB-binding protein

Zuwen Zhang and Beat Lutz*

Research Group Molecular Genetics of Behavior, Max-Planck-Institute of Psychiatry, Kraepelinstrasse 2–10, D-80804 Munich, Germany

*To whom correspondence should be addressed. Tel: +49 89 30 622 640; Fax: +49 89 30 622 642; Email: lutz{at}mpipsykl.mpg.de

CREB-binding protein (CBP) is a multifunctional cofactor implicated in many intracellular signal transduction pathways. We aimed to investigate the involvement of CBP in the cAMP response element-binding protein (CREB)-mediated pathway. The point mutation Tyr658Ala in the CREB-binding domain (CBD) was shown to abolish the binding activity of CBP to phospho-CREB, the activated form of CREB. By using a mutant Cre/loxP recombination system, this point mutation was aimed to be generated in the mouse genome in a tissue- and time-specific manner. A targeting construct in which CBD exon 5 and inverted exon 5* containing the point mutation flanked by two mutant loxP sites (lox66 and lox71) oriented in a head-to-head position was generated. When Cre recombinase is present, the DNA flanked by the two mutant loxP sites is inverted, forming one loxP and one double mutated loxP site. As the double mutated loxP site shows low affinity for Cre recombinase, the favorable reaction leads to a product where the mutated exon 5* is placed into the position to be correctly transcribed and spliced. Inversion was observed to be complete in both bacteria and mouse embryonic stem cells. Our results indicate that this Cre- mediated inversion method is a valuable tool to introduce point mutations in the mouse genome in a regulatable manner.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 JEMHome page
A. Jani, M. Wan, J. Zhang, K. Cui, J. Wu, P. Preston-Hurlburt, R. Khatri, K. Zhao, and T. Chi
A novel genetic strategy reveals unexpected roles of the Swi-Snf-like chromatin-remodeling BAF complex in thymocyte development
J. Exp. Med., November 24, 2008; 205(12): 2813 - 2825.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. R. Bayascas, K. Sakamoto, L. Armit, J. S. C. Arthur, and D. R. Alessi
Evaluation of Approaches to Generation of Tissue-specific Knock-in Mice
J. Biol. Chem., September 29, 2006; 281(39): 28772 - 28781.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
L. A. Shakes, D. M. Garland, D. K. Srivastava, K. R. Harewood, and P. K. Chatterjee
Minimal cross-recombination between wild-type and loxP511 sites in vivo facilitates truncating both ends of large DNA inserts in pBACe3.6 and related vectors
Nucleic Acids Res., August 1, 2005; 33(13): e118 - e118.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
P. Oberdoerffer, K. L. Otipoby, M. Maruyama, and K. Rajewsky
Unidirectional Cre-mediated genetic inversion in mice using the mutant loxP pair lox66/lox71
Nucleic Acids Res., November 15, 2003; 31(22): e140 - e140.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.