Cellectis - bioresearch

Designing the most optimized products for targeted integration

We are integrating a natural meganuclease recognition site in the most popular cell lines. This recognition site is part of an engineered “landing pad” called pi10.3^® that contains the genetic elements favoring an optimized clone selection strategy.

“Wild-type” cells are hygromycin resistant and contain a natural meganuclease recognition site. Upon expression of the corresponding natural meganuclease following cell transfection, a DNA double-strand break is made which triggers repair through homologous recombination. The transfected integration matrix is used for repair and contains upstream and downstream homology regions, a puromycin resistance gene, promoter and terminator sequences of the Gene Of Interest (GOI) coding sequence, and a truncated neomycin resistance gene. Upon repair, the GOI is integrated at the meganuclease recognition site, and the cells become puromycin and neomycin resistant.

NB : the genetic elements, the antibiotics used for selection and the selection protocol may vary from one cGPS^® cell line to another.