A pull-down assay using DNA/RNA-conjugated beads with a customized competition strategy: An effective approach to identify DNA/RNA binding proteins

Abstract

Innate immune response is insisted upon detection of foreign intracellular DNA or RNA derived from viruses and bacteria. This reaction is important to initiate an effective protective response for the host cells. This crucial step is induced by cytosolic nucleic acids sensors/binding proteins, which triggers the production of type I or type III interferons (IFNs) and proinflammatory cytokines such as Interleukin 6 (IL-6). The identification of these cytosolic DNA or RNA sensors is a key step in understanding the signaling pathways triggered by those pathogens. Here we describe an effective approach to identify potential known/novel DNA or RNA sensors: a pull-down assay using DNA/RNA-conjugated beads with a customized competition strategy, which conferred a more effective and efficient way to determine the interaction between DNA/RNA and the sensor protein(s), therefore greatly improves the progress to investigate potential novel cytosolic DNA or RNA sensors/ binding proteins • The customized method makes the traditional pull-down assays more effective and efficient to identify DNA/RNA binding protein(s). • With the competitor of your choice, the method provides specific information about the competitive binding between DNA/RNA and binding proteins.

Keywords

• A pull-down assay
• DNA/RNA-conjugated beads
• Competition
• Cytosolic DNA/RNA sensors