A split green fluorescent protein system to enhance spatial and temporal sensitivity of translating ribosome affinity purification.

ABSTRACT: Translating ribosome affinity purification (TRAP) utilizes transgenic plants expressing a ribosomal protein fused to a tag for affinity co-purification of ribosomes and the mRNAs that they are translating. This population of actively translated mRNAs (translatome) can be interrogated by quantitative PCR or RNA sequencing. Condition- or cell-specific promoters can be utilized to isolate the translatome of specific cell types, at different growth stages and/or in response to environmental variables. While advantageous for revealing differential expression, this approach may not provide sufficient sensitivity when activity of the condition/cell-specific promoter is weak, when ribosome turnover is low in the cells of interest, or when the targeted cells are ephemeral. In these situations, expr

SUBMITTER: Dinkeloo K 

PROVIDER: S-EPMC9544980 | biostudies-literature | 2022 Jul

REPOSITORIES: biostudies-literature