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Applications: Monitor ADAR1 activity.Study the effect of compounds on ADAR1 activity.
Background: ADAR (Adenosine Deaminase Acting on RNA) enzymes perform adenosine to inosine base editing in RNA, particularly targeting adenosines located within a specific double-stranded stem-loop motif (Figure 1). In the context of healthy, uninfected cells, ADAR1 performs A-to-I editing on endogenous double-stranded RNA to prevent it from activating the downstream dsRNA sensors RIG-I (retinoic acid-inducible gene I) and MDA5 (melanoma differentiation-associated protein 5), which in-turn activate a pro-inflammatory response. Loss of function mutations in ADAR1 result in aberrant activation of the dsRNA sensors and are involved in autoimmune disorders. ADAR1 dysfunction also impacts cancer cell growth, proliferation, and response to immunotherapy. ADAR1 expression is increased in many tumor types and ADAR1 knock-out has shown to improve the response to certain immunotherapies such PD-1 (programmed death protein 1)/PD-L1 (programmed death ligand 1) blockade and to circumvent tumor immunotherapy resistance mechanisms, making ADAR1 an attractive target for therapeutic development.
Description: ADAR1 Activity Luciferase Reporter HEK293 Cell Line is a HEK293 cell line designed to monitor the activity of ADAR1 (adenosine deaminase acting on RNA) enzyme. These cells were engineered to express ADAR1 (NM_001111.5) and an ADAR1 reporter construct comprised of an ADAR1 hairpin target with a stop codon (UAG) susceptible to ADAR1-mediated editing to tryptophan (UUG), located upstream of a firefly luciferase reporter.This cell line has been validated by comparing reporter activation to ADAR1 Responsive Luciferase Reporter HEK293 Cell Line (#82238) and in response to treatment with ADAR1 siRNA.The ADAR1 reporter construct is comprised of an ADAR1 hairpin target with a stop codon (UAG) upstream of the sequence encoding luciferase. In the absence of ADAR1, luciferase is not transcribed, and the cells show no luciferase activity. In the presence of ADAR1 activity, adenine is converted into inosine, encoding now the amino acid tryptophan (UUG) and enabling transcription and expression of luciferase. ADAR1 activity, therefore, directly correlates with luciferase activity.
Host Cell Line: HEK293
Mycoplasma Testing: The cell line has been screened to confirm the absence of Mycoplasma species.
Storage Stability: Cells will arrive upon 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. Contact technical support at support@bpsbioscience.com if the cells are not frozen in dry ice upon arrival.
Supplied As: Each vial contains >1 x 106 cells in 1 ml of Cell Freezing Medium (BPS Bioscience #79796
Uniprot: P55265
Warnings: Avoid freeze/thaw cycles
Biosafety Level: BSL-2
References: Bhate A., et al., 2019 Mol Cell. 73(5):866-868.
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Yuan, J., et al., 2023 J Exp Clin Cancer Res 42:149.
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