This manual describes how to use the Flp-In™ System to generate stable mammalian expression cell lines. The system uses Flp recombinase-mediated integration technology. The system includes Core System and Complete System. Core System includes vectors and primers for sequencing. Complete System includes Core System and selection agents.
The Flp-In System allows for the integration and expression of a gene of interest in mammalian cells at a specific genomic location. It utilizes a Flp Recombination Target (FRT) site inserted into the genome of a host cell line. An expression vector containing the gene of interest is then integrated into this specific site via Flp recombinase-mediated DNA recombination.
The system allows for the integration and expression of a gene of interest in mammalian cells at a specific genomic location.
The pFRT/ lacZeo plasmid is transfected into a mammalian cell line, and cells are selected for Zeocin™ resistance to identify clones with an integrated FRT site.
Stable expression cell lines are selected based on hygromycin resistance, Zeocin™ sensitivity, and a lack of β-galactosidase activity.