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Trap-TRAP, a Versatile Tool for Tissue-Specific Translatomics in Zebrafish

Developmental and physiological processes depend on the transcriptional and translational activity of heterogeneous cell populations. A main challenge in gene expression studies is dealing with this intrinsic complexity while keeping sequencing efficiency. Translating ribosome affinity purification...

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Autores principales: Corbacho, Jorge, Sanabria-Reinoso, Estefanía, Buono, Lorena, Fernández-Miñan, Ana, Martínez-Morales, Juan R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8841413/
https://www.ncbi.nlm.nih.gov/pubmed/35174174
http://dx.doi.org/10.3389/fcell.2021.817191
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author Corbacho, Jorge
Sanabria-Reinoso, Estefanía
Buono, Lorena
Fernández-Miñan, Ana
Martínez-Morales, Juan R.
author_facet Corbacho, Jorge
Sanabria-Reinoso, Estefanía
Buono, Lorena
Fernández-Miñan, Ana
Martínez-Morales, Juan R.
author_sort Corbacho, Jorge
collection PubMed
description Developmental and physiological processes depend on the transcriptional and translational activity of heterogeneous cell populations. A main challenge in gene expression studies is dealing with this intrinsic complexity while keeping sequencing efficiency. Translating ribosome affinity purification (TRAP) methods have allowed cell-specific recovery of polyribosome-associated RNAs by genetic tagging of ribosomes in selected cell populations. Here we combined the TRAP approach with adapted enhancer trap methods (trap-TRAP) to systematically generate zebrafish transgenic lines suitable for tissue-specific translatome interrogation. Through the random integration of a GFP-tagged version of the large subunit ribosomal protein L10a (EGFP-Rpl10a), we have generated stable lines driving expression in a variety of tissues, including the retina, skeletal muscle, lateral line primordia, rhombomeres, or jaws. To increase the range of applications, a UAS:TRAP transgenic line compatible with available Gal4 lines was also generated and tested. The resulting collection of lines and applications constitutes a resource for the zebrafish community in developmental genetics, organ physiology and disease modelling.
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spelling pubmed-88414132022-02-15 Trap-TRAP, a Versatile Tool for Tissue-Specific Translatomics in Zebrafish Corbacho, Jorge Sanabria-Reinoso, Estefanía Buono, Lorena Fernández-Miñan, Ana Martínez-Morales, Juan R. Front Cell Dev Biol Cell and Developmental Biology Developmental and physiological processes depend on the transcriptional and translational activity of heterogeneous cell populations. A main challenge in gene expression studies is dealing with this intrinsic complexity while keeping sequencing efficiency. Translating ribosome affinity purification (TRAP) methods have allowed cell-specific recovery of polyribosome-associated RNAs by genetic tagging of ribosomes in selected cell populations. Here we combined the TRAP approach with adapted enhancer trap methods (trap-TRAP) to systematically generate zebrafish transgenic lines suitable for tissue-specific translatome interrogation. Through the random integration of a GFP-tagged version of the large subunit ribosomal protein L10a (EGFP-Rpl10a), we have generated stable lines driving expression in a variety of tissues, including the retina, skeletal muscle, lateral line primordia, rhombomeres, or jaws. To increase the range of applications, a UAS:TRAP transgenic line compatible with available Gal4 lines was also generated and tested. The resulting collection of lines and applications constitutes a resource for the zebrafish community in developmental genetics, organ physiology and disease modelling. Frontiers Media S.A. 2022-01-31 /pmc/articles/PMC8841413/ /pubmed/35174174 http://dx.doi.org/10.3389/fcell.2021.817191 Text en Copyright © 2022 Corbacho, Sanabria-Reinoso, Buono, Fernández-Miñan and Martínez-Morales. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cell and Developmental Biology
Corbacho, Jorge
Sanabria-Reinoso, Estefanía
Buono, Lorena
Fernández-Miñan, Ana
Martínez-Morales, Juan R.
Trap-TRAP, a Versatile Tool for Tissue-Specific Translatomics in Zebrafish
title Trap-TRAP, a Versatile Tool for Tissue-Specific Translatomics in Zebrafish
title_full Trap-TRAP, a Versatile Tool for Tissue-Specific Translatomics in Zebrafish
title_fullStr Trap-TRAP, a Versatile Tool for Tissue-Specific Translatomics in Zebrafish
title_full_unstemmed Trap-TRAP, a Versatile Tool for Tissue-Specific Translatomics in Zebrafish
title_short Trap-TRAP, a Versatile Tool for Tissue-Specific Translatomics in Zebrafish
title_sort trap-trap, a versatile tool for tissue-specific translatomics in zebrafish
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8841413/
https://www.ncbi.nlm.nih.gov/pubmed/35174174
http://dx.doi.org/10.3389/fcell.2021.817191
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