Ransey, ElizabethBjörkbom, AndersLelyveld, VictorBiecek, PrzemyslawPantano, LorenaSzostak, JackSliz, Piotr2018-01-182017Ransey, Elizabeth, Anders Björkbom, Victor S. Lelyveld, Przemyslaw Biecek, Lorena Pantano, Jack W. Szostak, and Piotr Sliz. 2017. “Comparative analysis of LIN28-RNA binding sites identified at single nucleotide resolution.” RNA Biology 14 (12): 1756-1765. doi:10.1080/15476286.2017.1356566. http://dx.doi.org/10.1080/15476286.2017.1356566.http://nrs.harvard.edu/urn-3:HUL.InstRepos:34651993ABSTRACT It remains a formidable challenge to characterize the diverse complexes of RNA binding proteins and their targets. While crosslink and immunoprecipitation (CLIP) methods are powerful techniques that identify RNA targets on a global scale, the resolution and consistency of these methods is a matter of debate. Here we present a comparative analysis of LIN28-pre-let-7 UV-induced crosslinking using a tandem mass spectrometry (MS/MS) and deep sequencing interrogation of in vitro crosslinked complexes. Interestingly, analyses by the two methods diverge in their identification of crosslinked nucleotide identity – whereas bioinformatics and sequencing analyses suggest guanine in mammalian cells, MS/MS identifies uridine. This work suggests the need for comprehensive analysis and validation of crosslinking methodologies.en-USCLIPCIMSLIN28Journal Article2018-01-1810.1080/15476286.2017.1356566