A breakthrough in stable cell line generation!

cGPS^® targeted integration: easy and ready to use, saves time, effort and money.

The key benefits of meganuclease-driven targeted integration are:

• Little clone screening or the ability to work with bulk populations: high clone homogeneity is obtained thanks to very efficient targeted integration protocols (95%+ targeted integration),
• Guaranteed and predictable transgene expression: integration is performed in a single selected transcriptionally active genomic site,
• Highly stable expression: for at least 20 weeks, even in the absence of selection agents!
• Robust data comparison: the generated clones are isogenic,
• Fast: targeted cell population can be recovered in as little as 2 weeks, isolated clones in 3 to 4, due to the use of selection agents
• Easy: the use of a selection agent facilitates the recovery of targeted clones in the culture medium,
• Reproducible: highly efficient integration of a single transgene copy at a unique genomic site.

Generation of stably expressing cell lines by ‘traditional’ stable transfection protocols usually result in various random integration events in multiple copies (due to plasmid concatemerization) and at various genomic sites. This totally random process lead to high clone genotypic heterogeneity and uncontrolled expression levels. Besides, the level of expression of the randomly integrated transgenes is not stable overtime due to positional effects.

Site-specific recombination by use of recombinases is generally reversible and therefore leads to unstable gene expression.

cGPS and cGPS Custom products are immediately retargetable cell platforms. There is no prerequisite to first establish such a retargetable platform. Isogenic cell line generation is therefore very fast.

Targeted integration by way of meganuclease-driven homologous recombination comes with an easy two-step selection process. The use of selection agents:

• increases the efficiency of recovering clones with the desired phenotype (95%+ targeted integration efficiency after the use of selection agents),
• facilitates the recovery and analysis of targeted clones since they are directly added in in the culture medium,
• speeds up the process to targeted clone recovery.

cGPS^® and cGPS^® Custom reagents and protocols are the only products on the market that guarantee the fast and efficient selection of stable and isogenic clones with highly stable expression (even in the absence of selection agents).