Allyson Holmes1, Jean-Pierre Cabaniols1, Luc Selig1
1Cellectis bioresearch , 102 avenue Gaston Roussel, 93 235 Romainville Cedex, France
The development of cell-based assays for high-throughput screening (HTS) approaches often requires the generation of stable transformant cell lines. However, these cell lines are essentially created by random integration of a gene of interest (GOI) with no control over the level and stability of gene expression. We developed a targeted integration system in CHO cells, called cellular genome positioning system (cGPS®), based on the use stimulation of homologous gene targeting by meganucleases. Five different GOIs were knocked-in at the same locus in cGPS CHO-K1 cells. Further characterization revealed that the cGPS CHO-K1 system is more rapid (2-week protocol), efficient (all selected clones expressed the GOI), reproducible (GOI expression level variation of 12%), and stable over time (no change in GOI expression after 23 weeks of culture) than classical random integration. Moreover, in all cGPS CHO-K1 targeted clones, the recombinant protein was biologically active and it properties similar to the endogenous protein. This fast and robust method opens the door for creating large collections of cell lines expressing therapeutically relevant GOIs, enhancing the potential scope of HTS.
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