Color – Switch, stable reporter cell line CRE:
This cell line was transformed from the HEK293 cell line, which expresses the “LoxP-GFP-stop-LoxP-RFP” cassette under the CMV promoter (see Color-Switch cassette below). The cell line also contains an antibiotic blasticidin marker under the RSV promoter (not shown in the schematic).
- These CRE reporting cell lines are used to monitor the efficiency of CRE recombination in vivo. It is a great method and an easy tool to verify the recombination event by your CRE enzyme (your CRE expression plasmids, or pre-made CRE expression lentiviruses, or purified CRE protein) in vivo and in vitro.
- It is a convenient tool to verify your CRE-box-based system.
- It can also be used to verify CRISPR editing efficiency by targeting LoxP and the GFP / RFP region (note that we do not provide any sequence information unless a license is purchased for this cell line).
How does it work?
The cell line demonstrates a strong GFP (green fluorescent signal). Downstream RFP (red fluorescent protein) was not expressed due to the stop codon after GFP. Once the CRE protein is present in the nucleus, the CRE cleaves / removes the DNA fragment between two loxP sites, eliminating the stop codon (see DNA structure schematic above). As a result, the RFP is expressed and the cell line changes to fluorescent RFP. The GFP and RFP signal can be easily monitored through fluorescent cell sorting (for the ratio between GFP and RFP cells), or viewed with a fluorescent microscope, or fluorescent intensity measured if desired.
Sold at: 1 x vial (2 x 106 cells) / vial
Catalog No: SC018-Bsd