TRAP-SEQ of Eukaryotic Translatomes Applied to the Detection of Polysome-Associated Long Noncoding RNAs

TRAUBENIK, SOLEDAD; BLANCO, FLAVIO; ZANETTI, MARÍA EUGENIA; REYNOSO, MAURICIO

RNA tagging. Methods and Protocols

Springer

Año: 2019; p. 1 - 14

Translating ribosome affinity purification (TRAP) technology allows the isolation of polysomal complexesand the RNAs associated with at least one 80S ribosome. TRAP consists of the stabilization and affinitypurification of polysomes containing a tagged version of a ribosomal protein. Quantitative assessment ofthe TRAP RNA is achieved by direct sequencing (TRAP-SEQ), which provides accurate quantitation ofribosome-associated RNAs, including long noncoding RNAs (lncRNAs). Here we present an updatedprocedure for TRAP-SEQ, as well as a primary analysis guide for identification of ribosome-associatedlncRNAs. This methodology enables the study of dynamic association of lncRNAs by assessing rapidchanges in their transcript levels in polysomes at organ or cell-type level, during development, or inresponse to endogenous or exogenous stimuli.