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Updated: Jan 9, 2026
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Translating Ribosome Affinity Purification TRAP for RNA Isolation from Endothelial Cells In Vivo
Published on: May 25, 2019
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Analyzing the Translatome of Lymphatic and Venous Endothelial Cells In Vivo via Translating Ribosome Affinity
Olamide Olayinka1, Leia Zarinebaf1, Hyun Min Jung1
1Department of Pharmacology and Regenerative Medicine, University of Illinois at Chicago, Chicago, IL, USA.
Bio-Protocol
|December 11, 2025
Summary
Researchers optimized translating ribosome affinity purification (TRAP) in zebrafish to study endothelial cell gene expression. This method allows in vivo analysis of vascular and lymphatic biology without cell sorting, advancing research in the field.
Area of Science:
- Vascular Biology
- Lymphatic Biology
- Zebrafish Models
Background:
- Zebrafish are valuable for studying vascular and lymphatic systems due to transparency and genetic manipulability.
- Translating ribosome affinity purification (TRAP) is effective but underutilized in zebrafish.
- Previous TRAP applications in zebrafish required cell dissociation or sorting, limiting its utility.
Purpose of the Study:
- To develop and optimize an in vivo TRAP protocol for isolating ribosome-associated mRNAs from zebrafish endothelial cells.
- To enable specific analysis of actively translating transcripts in vascular and lymphatic endothelial cells without cell sorting.
- To establish a robust platform for studying endothelial biology in zebrafish.
Main Methods:
- Developed a novel transgenic zebrafish line expressing HA-tagged Rpl10a under the mrc1a promoter.
- Optimized an in vivo TRAP protocol for endothelial cells, avoiding dissociation or sorting.
- Performed differential expression analysis on enriched endothelial transcripts.
Main Results:
- Successfully isolated ribosome-associated mRNAs from endothelial cells in vivo.
- Demonstrated robust upregulation of key vascular and lymphatic genes (e.g., flt4, kdrl, lyve1b).
- Achieved high specificity in capturing the endothelial cell translatome.
Conclusions:
- The optimized TRAP protocol provides a powerful tool for in vivo endothelial translatome analysis in zebrafish.
- This method facilitates the study of endothelial gene expression under various perturbations.
- The approach is compatible with downstream applications like qPCR and RNA-seq for comprehensive analysis.
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