RFP/GFP Safe-Harbor HEK293 Cell Line

Recombinant stable HEK293 cell line constitutively expressing RFP (Red Fluorescent Protein) and GFP (Green Fluorescent Protein), which have been stably integrated into the AAVS1 safe harbor locus on chromosome 19. Expression of RFP is driven by a CMV promoter, whereas GFP is driven by an EF1A promoter. The combined DNA fragment (CMV-RFP-bGH Poly A-EF1A-GFP-T2A-Puro-SV40 Poly A) (Figure 1) was integrated at the AAVS1 safe harbor locus using CRISPR/Cas9 technology. Cells were cloned by limiting dilution to obtain a monoclonal population.  Without the adverse effects resulting from random integrations of RFP-GFP into the HEK293 genome, this cell line behaves like parental HEK293 cells.