WebProtein-RNA interactions (PRIs) control pivotal steps in RNA biogenesis, regulate multiple physiological and pathological cellular networks, and are emerging as important drug … To identify RNA–protein interactions systematically in living cells, we developed a method that measures the cellular interactions between any tagged test RNA and any tagged test protein. The principle of incPRINT is the transient dual expression of a MS2-tagged test RNA and a FLAG-tagged test protein in HEK293T … See more To establish incPRINT, we performed a series of small-scale experiments using a ∼1-kb conserved region of the lncRNA Xist called the A repeat, hereafter … See more To test the scalability of incPRINT for systematic identification of RNA–protein interactions, we interrogated our customized library of ~3000 FLAG-tagged human … See more Next, we sought to test if incPRINT can robustly identify proteins interacting with transcripts expressed at low endogenous levels. Identification of proteins … See more Because many lncRNAs function as modular scaffolds, enabling binding of specific RBPs to discrete RNA domains1,5, we sought to test if incPRINT allows the … See more
Northwestern Blot Analysis: Detecting RNA–Protein Interaction …
WebApr 14, 2024 · Protein-biomolecule interactions play pivotal roles in almost all biological processes. For a biomolecule of interest, the identification of the interacting protein(s) is essential. For this need, although many assays are available, highly robust and reliable methods are always desired. By combining a substrate-based proximity labeling activity … WebFeb 23, 2024 · This protocol describes a bacterial three-hybrid (B3H) assay, an in vivo system that reports on RNA–protein interactions and can be implemented in both forward and reverse genetic experiments. photo editor and frames
Cell Press: STAR Protocols
WebJul 26, 2007 · sensitive method to detect protein–nucleic acid interactions1–6.Itis based on the observation that the electrophoretic mobility of a protein–nucleic acid complex is typically less than that of the free nucleic acid (Fig. 1). The current, widely-used assay differs little from that originally described by Fried and Crothers7 and Garner WebElectrophoretic mobility shift assay (EMSA) is a sensitive technique useful in the identification and characterization of protein interactors with nucleic acids. This assay provides an efficient method to study DNA or RNA binding proteins and to identify nucleic acid substrates. The specific interac … WebThe proximity ligation assay (PLA) is an immune staining method that detects protein-protein interactions in fixed cells. We describe here RNA-PLA, a simple adaptation of this … photo editor and background remover