Novel recombination system using Cre recombinase alpha complementation

• Published in: Biotechnology letters Vol. 29; no. 9; pp. 1315 - 1322
• Main Authors: Seidi, Azadeh, Mie, Masayasu, Kobatake, Eiry
• Format: Journal Article
• Language: English
• Published: Dordrecht Dordrecht : Springer Netherlands 01.09.2007; Springer; Springer Nature B.V
• Subjects: Biological and medical sciences; Biotechnology; Cre recombinase; Cytomegalovirus; Escherichia coli - physiology; Escherichia coli Proteins - genetics; Escherichia coli Proteins - metabolism; Fundamental and applied biological sciences. Psychology; Genes; Green Fluorescent Proteins - genetics; Integrases - genetics; Promoter Regions, Genetic - genetics; Protein Engineering - methods; Protein transduction; Recombinant Proteins - biosynthesis; Short-term expression; Temporal control; Toxicity; α-Complementation; Recombination; Temporal control; α-Complementation .Cre recombinase; Complementation; Transduction; Gene expression; Protein transduction .Short-term expression; Protein
• DOI: 10.1007/s10529-007-9406-6
• ISSN: 0141-5492

A major limitation for the use of Cre recombinase is its toxicity and a lack of temporal control over its activity. We have developed a new recombination system using Cre recombinase α-complementation. Cre recombinase was divided and one fragment (β) was introduced into cells between two loxP sites with a CMV promoter in the upstream. The gene of interest (EGFP) was positioned just downstream of this construct. Cre recombinase activity was recovered by adding the other part of the molecule (α) to cells as a protein fragment, as evidenced by the expression of EGFP under the control of the CMV promoter. The activity of fragmented cre reached 68% of that of the wild type enzyme at 1 μM α-protein.