In vivo PAR-CLIP (viP-CLIP) of liver TIAL1 unveils targets regulating cholesterol synthesis and secretion

Abstract Here, we describe viP-CLIP ( in vivo PAR-CLIP) that is capable to identify endogenous RBP targets in mammalian tissues, thereby facilitating the functional analysis of RBP-regulatory networks in vivo . Briefly, mice are exposed to 4-Thiouridine that is incorporated into newly transcribed RNAs in the organism, enabling an efficient UV-crosslinking in multiple tissues crucial for a successful experiment. We report a comprehensive protocol of the critical steps to adapt the cell culture PAR-CLIP 1 protocol to the viP-CLIP protocol carried out in mice.