ADAR (Adenosine Deaminase Acting on RNA) is a family of enzymes that specifically deaminate adenosine residues to inosine in RNA molecules. This process is crucial for RNA editing, which can impact gene expression and protein function.

Recombinant ADAR Protein

Recombinant ADAR protein is produced through recombinant DNA technology, wherein the gene encoding ADAR is inserted into a suitable expression vector, transformed into a host organism (commonly E. coli or yeast), and then cultured to produce the protein.

Production Process

• Gene Cloning: The ADAR gene is amplified and cloned into an expression vector with a suitable promoter.
• Transformation: The vector is introduced into a host cell line (e.g., bacterial or yeast).
• Expression: The transformed cells are cultured under conditions that induce protein expression.
• Purification: The ADAR protein is isolated from the cell lysate using chromatography techniques (e.g., affinity, ion exchange).

Structure and Function

• Structure: Recombinant ADAR protein typically consists of multiple domains, including catalytic and RNA-binding domains. The protein's three-dimensional structure is essential for its function.
• Function: ADAR proteins specifically deaminate adenosine to inosine in double-stranded RNA (dsRNA). This modification can alter RNA stability, splicing, and translation.

Applications

• Functional Studies: Used in research to study RNA editing mechanisms and their implications.
• Therapeutics: Potential applications in gene therapy to correct RNA editing defects.
• Diagnostics: Can be used to analyze RNA editing patterns in various diseases.

Quality Control

• Purity: Ensured by SDS-PAGE, HPLC.
• Activity: Confirmed through RNA editing assays.
• Stability: Assessed under different storage conditions.

Recombinant ADAR proteins are valuable tools for understanding RNA biology and developing potential therapeutic strategies targeting RNA editing pathways.