A-to-I RNA Editing Reporter Plasmid

Adenosine-to-inosine (A-to-I) RNA editing is a process, catalysed by adenosine deaminases acting on RNA (ADARs), in which adenosine is converted into inosine by deamination is recognized by the cellular machinery as guanosine. This A-to-I editing is an essential post-transcriptional regulation in mammals that occurs in protein-coding regions of mRNAs as well as non-coding regions with inverted Alu repeats. The A-to-I editing may lead to non-synonymous amino acid mutations, RNA secondary structure alterations, pre-mRNA processing changes, and miRNA-mRNA redirection, thereby affecting multiple cellular bioprocesses and biofunctions. Deficiencies in A-to-I RNA editing is associated with human diseases and pathophysiology.

To efficiently monitor and quantify the editing activity, researchers have developed a bioluminescent reporter system based on the R/G editing site of the GluA2 transcript. The A-to-I editing reporter plasmid contains a modified R/G editing site of the GluA2 transcript in which an amber stop codon (UAG) is introduced. A Firefly luciferase (FLuc) and a Nanoluciferase (NLuc) are placed upstream and downstream of the editing site, respectively. FLuc is constitutively expressed as internal normalization. NLuc is conditionally expressed upon reporter-transcript editing. Without editing, only FLuc is expressed. While after editing, the stop codon (UAG) located between FLuc and NLuc is recoded into a tryptophan codon (UGG) result in a FLuc-NLuc fusion protein. The editing level could be measured as the ratio between luminescence from Nanoluciferase and Firefly luciferase. The reporter plasmid also contains antibiotic marker for stable selection in mammalian cells.

RGBiotech offers the A-to-I RNA editing reporter plasmid as well as the positive and negative control plasmids to assist your research in high-throughput screening of inhibitors and inducers of ADARs.

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