The ADAR1 Luciferase Reporter HEK293 cell line is designed to monitor RNA editing by Adenosine deaminase acting on RNA (ADAR1). This cell line stably expresses ADAR1 under the control of a CMV promoter and a separate ADAR editing reporter construct expressed under the control of another CMV promoter. The reporter contains the gene encoding firefly luciferase, which is constitutively expressed in the cells, upstream of the gene encoding the GluA2 ADAR substrate followed by the Renilla luciferase gene. The sequence corresponding to GluA2 has been modified to contain an amber stop codon (UAG). When edited by ADAR, this stop codon (UAG) will be changed to UIG (A to I edit), which is read as tryptophan (UGG) by the translation machinery. This edit allows translation to occur all the way to the end of the reporter mRNA and results in the expression of Renilla luciferase. Conversely, in the absence of ADAR1 activity, translation terminates at the stop codon and Renilla is not expressed. Reporter activity is read out as the Renilla Luciferase/Firefly luciferase ratio whereby inhibition of ADAR activity, and thus the UAG (stop) to UGG (tryptophan) conversion rate, will result in a dose-dependent decrease in the Renilla luciferase/Firefly luciferase ratio.
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
ADAR1 performs adenosine to inosine base editing in RNA, particularly targeting adenosines located within a specific stem-loop motif structure. It is proposed that ADARs evolved to provide additional diversity to the transcriptome and while the majority of ADAR editing events occur in non-coding RNAs, some, including the canonical GluA2 editing site, alter the amino acid sequence of coding proteins. ADAR1 plays a role in innate immunity by mitigating interferon signaling. Dysfunction of ADAR1 results in autoimmune disorders, and impacts cancer cell growth and proliferation as well as tumor response to immunotherapy. Since ADAR recognizes double-stranded RNA, it also functions to suppress or modify RNA viruses. Thus, it is implicated in viral evolution and in the emergence of viral variants such as SARS-CoV-2 variants.
Application
Screen or titrate inhibitors of ADAR1 activity
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.