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. 2020 Aug 7;10(8):1160. doi: 10.3390/biom10081160

Figure 1.

Figure 1

Overview of cross-linking strategies. Depending on the sample type and goal of the set-up, no cross-linking, chemical- and UV-induced cross-linking could be used. Cross-links are marked with a red cross. Formaldehyde cross-linking induces RNA–protein cross-links (both double-stranded as single-stranded), as well as protein-protein cross-links. In contrast, UV light (254 nm) only induces protein–single-stranded RNA molecule (ssRNA) cross-links. Substituted methylene blue will open up a double-stranded RNA (dsRNA) structure by intercalation, allowing dsRNA protein binders to be cross-linked using visible light. The combination of methylene blue (visible light) and UV light (254 nm) induces both ssRNA–protein and dsRNA–protein cross-links. PAR (photoactivatable ribonucleoside-enhanced) cross-linking (365 nm) is a variant of conventional UV cross-linking, increasing cross-linking efficiency due to the incorporation of a photoactivatable nucleoside analog. Figure adapted from Wheeler et al., (2018) [25].