Fig. 8: The NaP-TRAP method can be adapted to study the translation of multiple ORFs simultaneously in vivo. | Nature Communications

Fig. 8: The NaP-TRAP method can be adapted to study the translation of multiple ORFs simultaneously in vivo.

From: NaP-TRAP reveals the regulatory grammar in 5’UTR-mediated translation regulation during zebrafish development

Fig. 8

a NaP-TRAP is an accessible, versatile, and quantitative method that measures the translation of thousands of reporters simultaneously through the immunocapture of FLAG-tagged nascent peptides. b Through the over-expression of an HA-tagged RBP. NaP-TRAP can be employed in conjunction with an RNA immunoprecipitation experiment to measure the effect of RBP recruitment on translation. c–f NaP-TRAP quantifies translation in a frame-specific manner. Through the incorporation of additional epitope tags in frames 2,3 or ORFs outside of the main open reading frame, the NaP-TRAP method can be utilized to: (1) measure out-of-frame translation in the main ORF (c), (2) detect IRES sequences in an unbiased manner through the use of a bicistronic reporter (d), (3) identify frameshifting elements (e), and (4) quantify stop codon readthrough (f).

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