ADAR1 Activity TWO-Luciferase Reporter HEK293 Cell Line
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 (Figure 1).
This cell line has been validated by treatment with ADAR1 siRNA and the ADAR1 inhibitor Fludarabine.
Figure 1: Illustration of the mechanism of action of ADAR1 Activity TWO-Luciferase Reporter HEK293 Cell Line.
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.
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Materials Required for Cell Culture
| Name | Ordering Information |
| Thaw Medium 1 | BPS Bioscience #60187 |
| Growth Medium 1Y | BPS Bioscience #82535 |
Materials Required for Cellular Assays
| Name | Ordering Information |
| ADAR1 Responsive Luciferase Reporter HEK293 Cell Line | BPS Bioscience #82238 |
| ADAR1 Targeting siRNA | Horizon #M-008630-01-0005 |
| Lipofectamine RNAi Max | Thermo Fisher #13778030 |
| Assay Medium 1A | BPS Bioscience #79805 |
| Assay Medium: Thaw Medium 1 | BPS Bioscience #60187 |
| Fludarabine | BPS Bioscience #82534 |
| 96-well tissue culture white, clear-bottom assay plate | Corning #3610 |
| TWO-Step Luciferase (Firefly and Renilla) Assay System | BPS Bioscience #60690 |
| Luminometer |
The cell line has been screened to confirm the absence of Mycoplasma species.
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.
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.
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