title: Insect cell line development using FLP-mediated cassette exchange technology.
abstract: Traditional cell line development is quite laborious and time-consuming as it is based on the random integration of the gene of interest which leads to unpredictable expression behavior. In opposition, recombinase-mediated cassette exchange systems represent a powerful genetic engineering approach, allowing site-specific insertion of recombinant genes into pre-tagged genomic loci with superior expression characteristics, thus bypassing the need for extensive clone screening and shortening the development timelines. Such systems have not been widely implemented in insect cell lines used for the production of recombinant proteins most commonly through the baculovirus expression vector system. Herein, it is provided the protocol for the implementation of a FLP-mediated cassette exchange system in Spodoptera frugiperda Sf 9 cells, in order to grant a flexible cell line for the stable production of recombinant proteins.
authors.affiliation: Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal.
authors.forename: João
authors.initials: J
authors.lastname: Vidigal
authors.forename: Fabiana
authors.initials: F
authors.lastname: Fernandes
authors.forename: Ana S
authors.initials: AS
authors.lastname: Coroadinha
authors.forename: Ana P
authors.initials: AP
authors.lastname: Teixeira
authors.forename: Paula M
authors.initials: PM
authors.lastname: Alves
mesh_terms: D000818:Animals; D015139:Blotting, Southern; D002460:Cell Line; D004247:DNA / Q000302:isolation & purification; D004254:DNA Nucleotidyltransferases / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D007313:Insecta / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D061987:Sf9 Cells; D018411:Spodoptera / Q000166:cytology; D014162:Transfection
chemical_list: D004247:DNA; D004254:DNA Nucleotidyltransferases; C045119:FLP recombinase