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.
Mycoplasma testing: The cell line has been screened to confirm the absence of Mycoplasma species.
Host Species: human
Supplied as Each vial contains 2 x 106 cells in 1 ml of cell freezing medium (Biohippo #79796)
Background
AAVS1 (also known as the PPP1R12C locus) on human chromosome 19 is a well-validated 'safe harbor' site for hosting DNA transgenes. AAVS1 has an open chromatin structure and is transcription competent. Most importantly, disrupting the AAVS1 locus by inserting DNA transgenes has no known adverse effects on the cells. Specifically targeting the AAVS1 locus is a major advantage compared to the random integration obtained using other approaches such as lentivirus infection or cell transfection, which may cause insertional mutagenesis or disrupt important genes or cellular processes.
Application
Use as a control in cell-based assays.
Host Cell
HEK293, Human Embryonic Kidney, epithelial-like cells, adherent
Shipping Temperature
-80°C (dry ice)
Note
License Disclosure: Visit Biohippobioscience.com/license for the label license and other key information about this product. Troubleshooting Guide: Visit Biohippobioscience.com/cell-line-faq for detailed troubleshooting instructions. For all further questions, please email [email protected]. Warning: Avoid freeze/thaw cycles
Storage Stability
Cells are shipped in dry ice and should immediately be thawed or stored in liquid nitrogen upon receipt. Do not use a -80°C freezer for long term storage.