Invitrogen can create your cell line by transfection of a DNA plasmid and subsequent selection or by using lentivirus particles. Lentiviral particles allow very efficient DNA integration into the recipient cell genome and can reduce the timeline of creating the stable cell line.
The gene of interest or RNAi sequence will be randomly integrated into the cell’s genome. To find at least one clone that meets your expression requirements, we will expand and test many stable clones. The final stable cell line will be tested for mycoplasma and is supplied ready for further experiments.
Generation of Flp-In™ host cell lLinesFlp-In™ host cell lines have a single Flp recombinase target (FRT) site located at a transcriptionally active genomic locus that allows you to use efficiently generate isogenic expression cell lines using targeted integration. These host cell lines are generated by transfection with the Flp-In™ target site vector pFRT/lacZeo and selection with Zeocin™ agent. Transfected clones are screened by Southern blot and -galactosidase assay to identify those that have a single FRT integrated at a transcriptionally active locus. Once generated, the Flp-In™ host cell line can be used to generate Flp-In™ expression cell lines, each expressing a gene of interest at the same level. These cell lines are useful for comparing different genes of the same family, point mutations, splice variants and promoters.
Generation of Flp-In™ expression cell linesInvitrogen can generate isogenic expression cell lines by co-transfecting a Flp-In™ host cell line with the Flp-In™ expression vector and the Flp recombinase expression vector, pOG44. The transiently expressed recombinase efficiently integrates the expression vector at the genomic FRT site. As a result, all hygromycin B-resistant cells will be isogenic, with the expression vector integrated at the same locus. Since the Flp-In™ System creates isogenic cell lines, all transfected clones produce equivalent levels of the protein of interest. Invitrogen can also clone your gene of interest into a
Flp-In™ expression vector with our
custom cloning services.
T-REx™ cell line adaptation for regulated expressionTetracycline-Regulated Expression (
T-REx™) cell lines stably express the tetracycline (Tet) repressor and are designed for use with the T-REx™ System. The T-REx™ System is a tetracycline-regulated expression system that yields higher levels of induced expression than any other regulated mammalian expression system. This system can be used to regulate the expression of short hairpin RNAs (shRNAs), microRNA (miRNA) and gene sequences. If your cell line does not produce the tetracycline repressor protein to effectively utilize this system, we can adapt it so you can successfully use the T-REx™ system.
Your partner in discoveryIf you need to have your cell line stably express a protein or RNAi sequence, Invitrogen can help you through that. We have the experience and expertise to deliver the quality you deserve. To discuss your project, please contact Invitrogen Custom Services.
