An improved Flp deleter mouse in C57Bl/6 based on Flpo recombinase.
Recently, a codon improved version of the Flpe site specific recombinase, termed Flpo, was reported as having greatly improved performance in mammalian cell applications. However, the degree of improvement could not be estimated because essentially no Flpe activity was observed. Here, we compare Flpe and Flpo accurately in a mammalian cell assay to estimate that Flpo is about five times more active than Flpe and similar to Cre and Dre. Consequently, we generated a Flpo deleter mouse line from the JM8 C57Bl/6 ES cells used in the EUCOMM and KOMP systematic knock-out programs. In breeding experiments, we show that the Flpo deleter delivers complete recombination using alleles that are incompletely recombined by a commonly used Flpe deleter. This indicates that the Flpo deleter is more efficient.
Pubmed ID: 20506501 RIS Download
Animals | Base Sequence | Binding Sites | Cell Line | Cells, Cultured | DNA Nucleotidyltransferases | Embryo, Mammalian | Embryonic Stem Cells | Female | Fibroblasts | Gene Expression Regulation, Developmental | Gene Knockout Techniques | Green Fluorescent Proteins | Isoenzymes | Male | Mice | Mice, Inbred C57BL | Mice, Transgenic | Plasmids | Recombination, Genetic | Transfection