Products from BPS Bioscience require a minimum order value above 400€Applications: Use for gene knockout, transgene knock-in, mutagenesis, transgene integration, or other genome editing-related applications.
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.
Description: Recombinant stable HEK293 cell line constitutively expressing a FLAG-tagged Cas9 nuclease and GFP (Green Fluorescent Protein), which have been stably integrated into the AAVS1 safe harbor locus on chromosome 19. When transfected or transduced with single guide RNAs (sgRNAs), this cell line will sustain double-strand DNA breaks (DSBs) at targeted genome sites.Expression of Cas9 is driven by a CBh promoter, whereas GFP is driven by an EF1A promoter. The combined DNA fragment (CBh-Cas9-bGH Poly A-EF1A-GFP-T2A-Puro-SV40 Poly A), shown in Figure 1, was integrated at the AAVS1 safe harbor locus using CRISPR/Cas9 technology. The construct also contains a puromycin resistance gene. Cells were cloned by limiting dilution to obtain a monoclonal population. Without the adverse effects resulting from random integrations of Cas9/GFP into the HEK293 genome, this cell line behaves like parental HEK293 cells.
Figure 1: Transgene integration at the AAVS1 locus.
Host Cell Line: HEK293, Human Embryonic Kidney, epithelial-like cells, adherent
Mycoplasma Testing: The cell line has been screened to confirm the absence of Mycoplasma species.
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.
Supplied As: Each vial contains 2 x 10
6 cells in 1 ml of cell freezing medium (BPS Bioscience #79796)
Warnings: Avoid freeze/thaw cycles
Biosafety Level: BSL-2
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