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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...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
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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. |
format | Online Article Text |
id | pubmed-9606109 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
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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