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Heterologous reporter expression in the planarian Schmidtea mediterranea through somatic mRNA transfection

Planarians have long been studied for their regenerative abilities. Moving forward, tools for ectopic expression of non-native proteins will be of substantial value. Using a luminescent reporter to overcome the strong autofluorescence of planarian tissues, we demonstrate heterologous protein express...

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Autores principales: Hall, Richard Nelson, Weill, Uri, Drees, Leonard, Leal-Ortiz, Sergio, Li, Hongquan, Khariton, Margarita, Chai, Chew, Xue, Yuan, Rosental, Benyamin, Quake, Stephen R., Sánchez Alvarado, Alejandro, Melosh, Nicholas A., Fire, Andrew Z., Rink, Jochen C., Wang, Bo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9606109/
https://www.ncbi.nlm.nih.gov/pubmed/36313809
http://dx.doi.org/10.1016/j.crmeth.2022.100298
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author Hall, Richard Nelson
Weill, Uri
Drees, Leonard
Leal-Ortiz, Sergio
Li, Hongquan
Khariton, Margarita
Chai, Chew
Xue, Yuan
Rosental, Benyamin
Quake, Stephen R.
Sánchez Alvarado, Alejandro
Melosh, Nicholas A.
Fire, Andrew Z.
Rink, Jochen C.
Wang, Bo
author_facet Hall, Richard Nelson
Weill, Uri
Drees, Leonard
Leal-Ortiz, Sergio
Li, Hongquan
Khariton, Margarita
Chai, Chew
Xue, Yuan
Rosental, Benyamin
Quake, Stephen R.
Sánchez Alvarado, Alejandro
Melosh, Nicholas A.
Fire, Andrew Z.
Rink, Jochen C.
Wang, Bo
author_sort Hall, Richard Nelson
collection PubMed
description Planarians have long been studied for their regenerative abilities. Moving forward, tools for ectopic expression of non-native proteins will be of substantial value. Using a luminescent reporter to overcome the strong autofluorescence of planarian tissues, we demonstrate heterologous protein expression in planarian cells and live animals. Our approach is based on the introduction of mRNA through several nanotechnological and chemical transfection methods. We improve reporter expression by altering untranslated region (UTR) sequences and codon bias, facilitating the measurement of expression kinetics in both isolated cells and whole planarians using luminescence imaging. We also examine protein expression as a function of variations in the UTRs of delivered mRNA, demonstrating a framework to investigate gene regulation at the post-transcriptional level. Together, these advances expand the toolbox for the mechanistic analysis of planarian biology and establish a foundation for the development and expansion of transgenic techniques in this unique model system.
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spelling pubmed-96061092022-10-28 Heterologous reporter expression in the planarian Schmidtea mediterranea through somatic mRNA transfection Hall, Richard Nelson Weill, Uri Drees, Leonard Leal-Ortiz, Sergio Li, Hongquan Khariton, Margarita Chai, Chew Xue, Yuan Rosental, Benyamin Quake, Stephen R. Sánchez Alvarado, Alejandro Melosh, Nicholas A. Fire, Andrew Z. Rink, Jochen C. Wang, Bo Cell Rep Methods Article Planarians have long been studied for their regenerative abilities. Moving forward, tools for ectopic expression of non-native proteins will be of substantial value. Using a luminescent reporter to overcome the strong autofluorescence of planarian tissues, we demonstrate heterologous protein expression in planarian cells and live animals. Our approach is based on the introduction of mRNA through several nanotechnological and chemical transfection methods. We improve reporter expression by altering untranslated region (UTR) sequences and codon bias, facilitating the measurement of expression kinetics in both isolated cells and whole planarians using luminescence imaging. We also examine protein expression as a function of variations in the UTRs of delivered mRNA, demonstrating a framework to investigate gene regulation at the post-transcriptional level. Together, these advances expand the toolbox for the mechanistic analysis of planarian biology and establish a foundation for the development and expansion of transgenic techniques in this unique model system. Elsevier 2022-09-20 /pmc/articles/PMC9606109/ /pubmed/36313809 http://dx.doi.org/10.1016/j.crmeth.2022.100298 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Hall, Richard Nelson
Weill, Uri
Drees, Leonard
Leal-Ortiz, Sergio
Li, Hongquan
Khariton, Margarita
Chai, Chew
Xue, Yuan
Rosental, Benyamin
Quake, Stephen R.
Sánchez Alvarado, Alejandro
Melosh, Nicholas A.
Fire, Andrew Z.
Rink, Jochen C.
Wang, Bo
Heterologous reporter expression in the planarian Schmidtea mediterranea through somatic mRNA transfection
title Heterologous reporter expression in the planarian Schmidtea mediterranea through somatic mRNA transfection
title_full Heterologous reporter expression in the planarian Schmidtea mediterranea through somatic mRNA transfection
title_fullStr Heterologous reporter expression in the planarian Schmidtea mediterranea through somatic mRNA transfection
title_full_unstemmed Heterologous reporter expression in the planarian Schmidtea mediterranea through somatic mRNA transfection
title_short Heterologous reporter expression in the planarian Schmidtea mediterranea through somatic mRNA transfection
title_sort heterologous reporter expression in the planarian schmidtea mediterranea through somatic mrna transfection
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9606109/
https://www.ncbi.nlm.nih.gov/pubmed/36313809
http://dx.doi.org/10.1016/j.crmeth.2022.100298
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