Products from BPS Bioscience require a minimum order value above 400€
Applications: Monitor ADAR1 activity.Study the effect of compounds on ADAR1 activity.Simultaneous measurement of compound effect on ADAR1 activity and impact on cell viability.
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 been demonstrated 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 TWO-Luciferase Reporter HEK293 Cell Line is a HEK293 cell line designed to monitor cell viability in parallel with ADAR1 (adenosine deaminase acting on RNA) enzyme activity. These cells were engineered to express ADAR1 (NM_001111.5) and an ADAR1 reporter construct. This construct is 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. In addition, they constitutively express Renilla Luciferase under the control of a CMV promoter, which can be used to determine cell viability.This cell line has been validated by treatment with ADAR1 siRNA and the ADAR1 inhibitor Fludarabine.The ADAR1 reporter construct is comprised of an ADAR1 hairpin target with a stop codon (UAG) upstream of the sequence encoding firefly luciferase. In the absence of ADAR1, firefly luciferase is not transcribed, and the cells show no firefly luciferase activity. However, these cells were engineered to express ADAR1, which converts adenine into inosine, now encoding the amino acid tryptophan (UUG) and enabling transcription and expression of firefly luciferase. ADAR1 activity, therefore, directly correlates with firefly luciferase activity. Renilla luciferase is constitutively expressed from a separate construct and therefore Renilla luciferase activity correlates with the number of cells, but not with ADAR1 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. 2019 Mar 7;73(5):866-868.
Buchumenski I., et al., 2019 Nature 565(7737):43-48.
Gallo A., et al., 2017 Human Genetics 136(9): 1265-1278.
Ishizuka J.J., et al., 2018 Genes (Basel). 10(1):12.
Yuan J., et al., 2023 J Exp Clin Cancer Res 42: 149.
Zhang T., et al., 2022 Nature 606: 594-602.
English
Deutsch
