Pseudouridine counts among the most abundant modifications in cellular RNA, but its function so far remains elusive. Haiqi Xu, Linzhen Kong and collaborating researchers within the Song team, the Lu team and the Kriaucionis team at Ludwig Oxford, have introduced 2-bromoacrylamide-assisted cyclisation sequencing (BACS) to quantitatively profile pseudouridine at single-base resolution.
BACS is based upon novel bromoacrylamide cyclisation chemistry which allows the pseudouridine-to-cytosine transition, rather than truncating or deleting, which allows accurate quantification of pseudouridine stoichiometry and sequencing of pseudouridine at absolute single-base resolution. The group have used this new method to successfully detect all known pseudouridine sites in human ribosomal RNA and generated the first quantitative pseudouridine map of human small nucleolar RNA and transfer RNA.
They have subsequently applied BACS to elucidate the targets and sequence motifs of three key pseudouridine synthases in the HeLa cancer cell line and identified further abundant pseudouridine sites in Epstein-Barr virus encoded small RNAs. They anticipate the development of BACS to serve as a powerful tool to uncover the biological importance of pseudouridine in future studies.
Associate Professor Chunxiao Song, the principal investigator for this work, commented “I am delighted to see this methodology out in the public domain, and hope to see BACS play a key role as the universal standard for determining the function of pseudouridine in diverse biological processes”.
To find out more, see the full article in Nature Methods.