Antibody Barcode CLIP (ABC)
Unprecedented scalable mapping of the RBP-RNA interactome
RNA binding proteins (RBPs) influence cell state by modulating the RNA life cycle at several steps, including splicing, localization, translation and degradation. Insights into how RBPs affect RNA processing can be gleaned from the application of cross-linking, immunoprecipitation followed by sequencing (CLIP-seq/HITS-CLIP) based technologies. Despite recent advances in improving CLIP library generation (eCLIP/irCLIP), CLIP workflows are still time and labor-intensive while being limited to a single RBP target.
To improve the scalability and ease-of-use, we developed Antibody Barcode CLIP (ABC). This new technology leverages proximity ligations between barcoded antibodies and RNA bound by RBPs, followed by eCLIP library preparation to identify the binding sites of multiple RPBs simultaneously.