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SCREENflex
SCREENflex | GPCRs | Go to ... |
SCREENflex | Customized cell lines | Go to ... |
Overview
Conventional strategies to establish stable cell lines for primary screening are based on random integration of the desired expression-vectors into the host cells genome. Both, the number of integrated vectors as well as the site of integration tremendously influence the stability and level of drug target gene expression. Therefore, the isolation of a proper cell line is an extensive procedure without guaranteed success.
The SCREENflex technology is the result of more than 10 years of research and development and the hypothesis, that the expression of a single copy of any heterologous DNA is sufficient – as far as an appropriate locus has been identified.
Our SCREENflex master cell lines harbour such single tagged chromosomal loci guaranteeing stable expression of a transgene. A tagging and screening procedure has been developed to isolate a set of master cell lines which support gene expression levels that are optimal for cells for drug screening campaigns.
In a single transfection step, the tagging cassette of a SCREENflex master cell line is exchanged for a drug target gene or a customer's gene (that can be a GPCR, Ion channel or kinase) by a DNA-recombinase and a selection trap. Since no other chromosomal regions are touched, all positive expression and growth features of the optimized master cell line are transferred to the new screening cell line.
InSCREENeX master cell lines are based on CHO-K1 and HEK293 cells.
Our SCREENflex technology has been described in several peer-reviewed papers:
Rapid establishment of g-protein-coupled receptor-expressing cell lines by site-specific integration.Schucht R, Lydford S, Andzinski L, Zauers J, Cooper J, Hauser H, Wirth D, May T. J Biomol Screen. 2011 | link |
Integrated strategy for the production of therapeutic retroviral vectors.Carrondo M, Panet A, Wirth D, Coroadinha AS, Cruz P, Falk H, Schucht R, Dupont F, Geny-Fiamma C, Merten OW, Hauser H.Hum Gene Ther. 2011
Recombinant protein expression by targeting pre-selected chromosomal loci. Nehlsen K, Schucht R, da Gama-Norton L, Krömer W, Baer A, Cayli A, Hauser H, Wirth D. BMC Biotechnol. 2009
Precise regulation of transgene expression level and control of cell physiology. Schucht R, Wirth D, May T. Cell Biol Toxicol. 2009
A new generation of retroviral producer cells: predictable and stable virus production by Flp-mediated site-specific integration of retroviral vectors. Schucht R, Coroadinha AS, Zanta-Boussif MA, Verhoeyen E, Carrondo MJ, Hauser H, Wirth D. Mol Ther. 2006
GPCRs
G protein coupled receptors (GPCRs) are targets of the majority of current pharmaceuticals. Based on our SCREENflex master cell lines we have generated robust cell lines stably expressing common GPCRs. Please find here an application protocol, established by Molecular Devices (UK) using our SCREENflex-CHRM3 cell line. | Download |
Due to the unique development procedure, the cells remain stable for months without any selection pressure. The cell lines are distributed by ECACC (UK). | Click for catalogue |
Customized cell lines
Our SCREENflex technology allows a high level of flexibility. According to your need we establish stable cell lines expressing your desired target. | Click for request |