Document Detail


S Phase-preferential Cre-recombination in mammalian cells revealed by HIV-TAT-PTD-mediated protein transduction.
MedLine Citation:
PMID:  17965427     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The Cre recombinase of bacteriophage P1 is a powerful tool for artificial modification of genomic function in mammalian cells. To date, many researchers have studied the enzymatic biochemistry of Cre recombinase in loxP site-specific cleavage and rearrangement, as well as its use in gene technology. However, the intricate mechanisms of Cre-mediated recombination are still poorly understood. For example, more knowledge is needed in order to understand Cre recombinase's dependency on cell cycle, the necessity of other factors for recombination, and the exact nuclear environment that's required at the target locus, in order for recombination to take place in eukaryotic cells. In this study, we showed that P1 Cre-mediated recombination occurred frequently during S-phase of the cell cycle. HeLa cells were synchronized in cell cycle with the thymidine-hydroxyurea block method, and recombinant Cre proteins were fused with HIV-1 TAT protein transduction domains (PTD) in every phase of the cell cycle. Results showed that the transduction of PTD-Cre gave rise to genomic recombination preferentially during the S-phase of cell cycle. These findings will contribute significantly to the development of the Cre/loxP recombination system in vivo.
Authors:
Mayumi Hashimoto; Makoto Taniguchi; Susumu Yoshino; Shiho Arai; Kenzo Sato
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2007-10-27
Journal Detail:
Title:  Journal of biochemistry     Volume:  143     ISSN:  0021-924X     ISO Abbreviation:  J. Biochem.     Publication Date:  2008 Jan 
Date Detail:
Created Date:  2008-01-21     Completed Date:  2008-07-01     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0376600     Medline TA:  J Biochem     Country:  Japan    
Other Details:
Languages:  eng     Pagination:  87-95     Citation Subset:  IM    
Affiliation:
Division of Molecular Biology, School of Life Science, Faculty of Medicine, Tottori University, 86 Nishi-cho, Yonago 683-8503, Japan.
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MeSH Terms
Descriptor/Qualifier:
Cell Cycle / genetics
Genes, Reporter
Hela Cells
Humans
Integrases / genetics,  metabolism*
Protein Structure, Tertiary
Protein Transport
Recombinant Fusion Proteins / isolation & purification,  metabolism
Recombination, Genetic*
S Phase / genetics*
tat Gene Products, Human Immunodeficiency Virus / chemistry*,  genetics
Chemical
Reg. No./Substance:
0/Recombinant Fusion Proteins; 0/tat Gene Products, Human Immunodeficiency Virus; EC 2.7.7.-/Cre recombinase; EC 2.7.7.-/Integrases

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