biostudies-literatureXie JJapan Agency for Medical Research and DevelopmentTakeda Science Foundation; and performed as research programmes of the Grant-in-Aid for Scientific Research, the Japan Society for the Promotion of Science251-265https://www.ebi.ac.uk/biostudies/studies/S-EPMC12480733biostudies-literatureUnknown178(4)Long non-coding RNAs (lncRNAs) regulate a wide array of cellular processes through interactions with RNA-binding proteins (RBPs). Taurine Upregulated Gene 1 (TUG1) is an lncRNA that is overexpressed in many types of cancer and has been implicated in resolving R-loops, thereby maintaining genomic integrity. However, the full spectrum of its protein interactions and stress-responsive dynamics remains unclear. Here, we employed CRISPR-assisted RNA-protein interaction detection (CARPID) combined with mass spectrometry to comprehensively identify the interacting proteins of TUG1 in HEK293T cells. Using three distinct single-guide RNAs (sgRNAs) targeting different regions of TUG1, we consistently identified 17 TUG1-interacting proteins under basal conditions. Upon camptothecin (CPT) treatment, wJournal of biochemistryComprehensive identification of proteins interacting with long non-coding RNA TUG1 in R-loop regulation.PMC1248073323H0274723ck0106816h0001 and 23ama221204h000223K06634Watanabe SIto TKondo YNakagawa RXie JSuzuki MMShinjo KIijima KNishimura TfalseComprehensive identification of proteins interacting with long non-coding RNA TUG1 in R-loop regulation.Long non-coding RNAs (lncRNAs) regulate a wide array of cellular processes through interactions with RNA-binding proteins (RBPs). Taurine Upregulated Gene 1 (TUG1) is an lncRNA that is overexpressed in many types of cancer and has been implicated in resolving R-loops, thereby maintaining genomic integrity. However, the full spectrum of its protein interactions and stress-responsive dynamics remains unclear. Here, we employed CRISPR-assisted RNA-protein interaction detection (CARPID) combined with mass spectrometry to comprehensively identify the interacting proteins of TUG1 in HEK293T cells. Using three distinct single-guide RNAs (sgRNAs) targeting different regions of TUG1, we consistently identified 17 TUG1-interacting proteins under basal conditions. Upon camptothecin (CPT) treatment, w2025-01-01T00:00:00Z2025 Sep2026-05-31T03:12:55.091Z2026-05-31T03:09:23.374ZS-EPMC124807334065430610.1093/jb/mvaf042