NaP-TRAP reveals the regulatory grammar in 5'UTR-mediated translation regulation during zebrafish development.

ABSTRACT: The cis-regulatory elements encoded in an mRNA determine its stability and translational output. While there has been a considerable effort to understand the factors driving mRNA stability, the regulatory frameworks governing translational control remain more elusive. We have developed a novel massively parallel reporter assay (MPRA) to measure mRNA translation, named Nascent Peptide Translating Ribosome Affinity Purification (NaP-TRAP). NaP-TRAP measures translation in a frame-specific manner through the immunocapture of epitope tagged nascent peptides of reporter mRNAs. We benchmark NaP-TRAP to polysome profiling and use it to quantify Kozak strength and the regulatory landscapes of 5' UTRs in the developing zebrafish embryo and in human cells. Through this approach we identified general

SUBMITTER: Strayer EC 

PROVIDER: S-EPMC11685710 | biostudies-literature | 2024 Dec

REPOSITORIES: biostudies-literature