Fig. 1: agoTRIBE fuses AGO2 and the editing ___domain of ADAR2.
From: Detection of transcriptome-wide microRNA–target interactions in single cells with agoTRIBE
a, Schematic representation of the agoTRIBE method. The agoTRIBE approach fuses human Argonaute2 with the adenosine deaminase ___domain of human ADAR2, carrying a hyperactive mutation (E488Q) and depositing edits on the targeted transcripts. The edited nucleotides can be detected as A>G substitutions by either standard or single-cell RNA-seq. Top: schematic representation of the N-terminal tagging of human Argonaute2. b, Human Argonaute2 protein structure prediction using AlphaFold2. Tagging of the C terminus, which is embedded in the protein structure, could result in a misfolded protein unable to load miRNAs. c, Immunofluorescence staining to visualize agoTRIBE. AGO2 (green) and ADAR2 (red) immunofluorescence costaining (co-IF) were used to detect agoTRIBE, while DAPI (magenta) was used for nuclear staining. Each microscopy experiment was performed in at least two replicates and a representative experiment is shown. Colocalization in cytoplasmic foci suggests that agoTRIBE is present in P-bodies.
