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. 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.