ADAR1 Responsive Luciferase Reporter HEK293 Cell Line

Catalog #
82238
$6,500 *
Size: 2 vials
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Description

ADAR1 Responsive Luciferase Reporter HEK293 Cell Line is a HEK293 cell line designed to respond to ADAR1 enzymatic activity. These cells were engineered to express 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 (Figure 1).

This cell line has been validated by comparing reporter activation after transfection with ADAR1 and ADAR2. This cell line has been used for the development of the constitutive ADAR1 expressing ADAR1 Activity Luciferase Reporter HEK293 Cell Line (#82239).

Figure 1: Illustration of the mechanism of action of ADAR1 Responsive Luciferase Reporter HEK293 Cell Line.
The ADAR1 reporter construct is compromised of an ADAR1 hairpin targer with a stop codon (UAG) upstream of the sequence encoding luciferase. In the presence of ADAR1 activity, as in the case of transfection with ADAR1, adenine is converted into inosine encoding now the amino acid tryptophan (UUG) and enabling transcription and expression of luciferase. In the absence of transfected ADAR1, luciferase is not transcribed, and the cells show background luciferase activity. Luciferase activity directly correlated with ADAR1 activity.

Interested in screening and profiling ADAR mutants without the need to purchase and license the cell line? Check out our Cell Signaling Pathway Screening and Profiling Service.

Purchase of this cell line is for research purposes only; commercial use requires a separate license. View the full terms and conditions.

Synonyms
DRADA, IFI4, P136, DSH, AGS6, G1P1, DSRAD, IFI-4, DSRAD, ADAR-1
Product Info
Storage and Usage
Citations
Host Cell Line
HEK293, Human Embryonic Kidney, epithelial-like cells, adherent
Species
Human
Supplied As
Each vial contains >1 x 106 cells in 1 ml of Cell Freezing Medium (BPS Bioscience #79796)
Materials Required But Not Supplied

Materials Required for Cell Culture

Name Ordering Information
Thaw Medium 1 BPS Bioscience #60187
Growth Medium 1N BPS Bioscience #79801

 

Materials Required for Cellular Assays

Name Ordering Information
ADAR1 Activity Luciferase Reporter HEK293 Cell Line BPS Bioscience #82239
Plasmids encoding ADAR1 isoforms and ADAR2  
Lipofectamine™ 2000 Transfection Reagent Thermo Fisher #11668027
Assay Media: Assay Medium 1A BPS Bioscience #79805
Thaw Medium 1 BPS Bioscience #60187
96-well tissue culture white, clear-bottom assay plate Corning #3610
ONE-Step™ Luciferase Assay System BPS Bioscience #60690
Luminometer  
UniProt #
P55265
Mycoplasma Testing

The cell line has been screened to confirm the absence of Mycoplasma species.

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.

HEK293 cells express low levels of endogenous ADAR1, therefore this cell line is ideal for studies of genetically engineering variants of ADAR1. For example, one can compare the activity of ADAR1 variants introduced into the cells by transfection or transduction. The use of luciferase as reporter allows for an easy assay read-out, making this cell line an attractive cellular system for ADAR1 studies.

References

Bhate A., Sun T. and Li J.B., 2019 Mol Cell. 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., Manguso RT, Cheruiyot CK, Bi K, Panda A, Iracheta-Vellve A, Miller BC, Du PP, Yates KB, Dubrot J,
Xu L.D. and Öhman M., 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.