CHROMOSOME-DAMAGE INDUCED BY COMBINED TREATMENTS WITH RESTRICTION ENDONUCLEASES INTRODUCED INTO CHO CELLS BY SINGLE OR DOUBLE ELECTROPORATION

The possible recombination between non-homologous termini produced by restriction enzymes (REs) introduced in CHO cells by electroporation was studied. For this purpose, different combinations of REs that produced blunt or 5' overhanging DNA double-strand breaks were electroporated into cells either at the same time or separately by double electroporation experiments. Prior to double electroporation, it was confirmed that, once the cells have been electroporated, they resist a second electroporation, as assessed by cell viability analysis. Besides, the efficient and homogeneous introduction of labelled, non-permeable molecules was assessed by fluorescence microscopy. Our results showed interaction for most of the conditions, mainly when the REs were introduced separately. Differences found in the degree of interaction between the combinations studied are discussed.