Conditional mutagenesis by cell-permeable proteins: potential, limitations and prospects.
The combination of two powerful technologies, the Cre/loxP recombination system and the protein transduction technique, holds great promise for the advancement of biomedical and genome research by enabling precise and rapid control over mutation events. Protein transduction is a recently developed technology to deliver biologically active proteins directly into mammalian cells. It involves the generation of fusion proteins consisting of the cargo molecule to be delivered and a so-called protein transduction domain. Recently, the derivation of cell permeable variants of the DNA recombinase Cre has been reported. Cre is a site-specific recombinase that recognizes 34 base pair loxP sites and has been widely used to genetically engineer mammalian cells in vitro and in vivo. Recombinant cell-permeable Cre recombinase was found to efficiently induce recombination of loxP-modified alleles in various mammalian cell lines. Here we review recent advances in conditional expression and mutagenesis employing cell-permeable Cre proteins. Moreover, this review summarizes recent findings of studies aimed at deciphering the molecular mechanism of the cellular uptake of cell-permeable fusion proteins.
Patsch
C
C
Stem Cell Engineering Group, Institute of Reconstructive Neurobiology, University of Bonn, Life and Brain Center and Hertie Foundation, Sigmund-Freud Strasse 25, 53105 Bonn, Germany.
Edenhofer
F
F
D000595:Amino Acid Sequence; D000818:Animals; D002463:Cell Membrane Permeability; D004254:DNA Nucleotidyltransferases / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000235:genetics*; D017434:Protein Structure, Tertiary; D014161:Transduction, Genetic
D004254:DNA Nucleotidyltransferases; C077570:Site-specific recombinase