The Journal of Biochemistry
Online ISSN : 1756-2651
Print ISSN : 0021-924X
Efficiency of Recombination by Cre Transient Expression in Embryonic Stem Cells: Comparison of Various Promoters
Kimi ArakiTakashi ImaizumiKeiji OkuyamaYuichi OikeKen-ichi Yamamura
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JOURNAL FREE ACCESS

1997 Volume 122 Issue 5 Pages 977-982

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Abstract

The Cre-loxP recombination system of bacteriophage P1 is frequently utilized in genetic manipulation in embryonic stem (ES) cells. The level of Cre expression is critical to induce loxP site-specific recombination in ES cells. To compare the efficiency of recombination, we constructed four Cre expression vectors driven by different promoters: cytomegarovirus/chicken β-actin (CAG) promoter, human polypeptide chain elongation factor 1α (hEF-1α) promoter, mouse phosphoglycerate kinase-1 (mPGK) promoter, and polyoma enhancer/herpes simplex virus thymidine kinase (MC1) promoter. We introduced these Cre expression vectors by electroporation into three ES cell lines carrying a single copy of CAG-loxP-chloramphenicol acetyltransferase (CAT) gene-loxP-β-galactosidase (β-gal) gene construct. Since the Cre-mediated recombination leads to excision of the CAT gene, the efficiency of recombination can be monitored as β-gal expression. No selection system was used in the experiments. The maximum recombination frequency was obtained when the CAG promoter was used, followed by the hEF-1α promoter, the mPGK promoter and the MC1 promoter in order. These results indicate that the efficiency of recombination in transient expression system correlates with the promoter activity of Cre expression vector. Thus, it is important to choose the promoter for effective recombination by Cre.

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