Fig. 1
From: In-cell identification and measurement of RNA-protein interactions
Principle of the incPRINT method. HEK293T cells, stably expressing a NanoLuc luciferase-MS2CP recombinant protein from an integrated plasmid, were co-transfected with plasmids encoding a MS2-tagged test RNA and 3xFLAG-tagged test proteins in a 96-well format (each well contains cells expressing one tagged test RNA and one tagged test protein). Cell lysates were applied to anti-FLAG-coated 384-well plates to immuno-purify test proteins with their interacting RNAs. After washing off nonspecific interactions, the MS2-RNA/FLAG-tagged protein complexes were detected by NanoLuc luciferase, tethered to the test RNA through the MS2-MS2CP interaction. The expression levels of the FLAG-tagged test proteins were detected by ELISA using a second anti-FLAG antibody coupled to horseradish peroxidase (HRP).
