Streptavidin mediated CLEAR-CLIP in desmoplakin-BirA overexpressing mouse keratinocyte
Argonautes were found to be located at desomosal plaque. To specifically identify Argonaute binding microRNA-RNA species, Desomoplakin fused biotin ligase BirA was overexpressed in mouse keratinocytes. By pulling down Biotin labled proteins using streptavidin, we examines the functions of Argonautes at desmosomal plaques. Overall design: Desomoplakin fused with biotin ligase BirA was overexpressed in mouse keratinocytes. Streptavidin was used to pull down all biotinated proteins at the desmosomal plaque, including Argonautes. By sequencing microRNA-mRNA chimeras using CLEAR-CLIP technique, the function of Argonautes at desmosomal plaque could be inferred.