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In vivo modulation of endogenous gene expression via CRISPR/Cas9-mediated 3’UTR editing

The 3' untranslated regions (UTRs) modulate gene expression levels by regulating mRNA stability and translation. We previously showed that the replacement of the negative regulatory elements from the 3'UTR of glial cell line-derived neurotrophic factor (GDNF) resulted in increased endogeno...

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Autores principales: Mätlik, Kärt, Olfat, Soophie, Cowlishaw, Mark Cary, Moreno, Eva Domenech, Ollila, Saara, Andressoo, Jaan-Olle
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10009458/
https://www.ncbi.nlm.nih.gov/pubmed/36923835
http://dx.doi.org/10.1016/j.heliyon.2023.e13844
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author Mätlik, Kärt
Olfat, Soophie
Cowlishaw, Mark Cary
Moreno, Eva Domenech
Ollila, Saara
Andressoo, Jaan-Olle
author_facet Mätlik, Kärt
Olfat, Soophie
Cowlishaw, Mark Cary
Moreno, Eva Domenech
Ollila, Saara
Andressoo, Jaan-Olle
author_sort Mätlik, Kärt
collection PubMed
description The 3' untranslated regions (UTRs) modulate gene expression levels by regulating mRNA stability and translation. We previously showed that the replacement of the negative regulatory elements from the 3'UTR of glial cell line-derived neurotrophic factor (GDNF) resulted in increased endogenous GDNF expression while retaining its normal spatiotemporal expression pattern. Here, we have developed a methodology for the generation of in vivo hyper- and hypomorphic alleles via 3'UTR targeting using the CRISPR/Cas9 system. We demonstrate that CRISPR/Cas9-mediated excision of a long inhibitory sequence from Gdnf native 3'UTR in mouse zygotes increases the levels of endogenous GDNF with similar phenotypic alterations in embryonic kidney development as we described in GDNF constitutive and conditional hypermorphic mice. Furthermore, we show that CRISPR/Cas9-mediated targeting of 3’UTRs in vivo allows the modulation of the expression levels of two other morphogens, Gdf11 and Bdnf. Together, our work demonstrates the power of in vivo 3’UTR editing using the CRISPR/Cas9 system to create hyper- and hypomorphic alleles, suggesting wide applicability in studies on gene function and potentially, in gene therapy.
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spelling pubmed-100094582023-03-14 In vivo modulation of endogenous gene expression via CRISPR/Cas9-mediated 3’UTR editing Mätlik, Kärt Olfat, Soophie Cowlishaw, Mark Cary Moreno, Eva Domenech Ollila, Saara Andressoo, Jaan-Olle Heliyon Research Article The 3' untranslated regions (UTRs) modulate gene expression levels by regulating mRNA stability and translation. We previously showed that the replacement of the negative regulatory elements from the 3'UTR of glial cell line-derived neurotrophic factor (GDNF) resulted in increased endogenous GDNF expression while retaining its normal spatiotemporal expression pattern. Here, we have developed a methodology for the generation of in vivo hyper- and hypomorphic alleles via 3'UTR targeting using the CRISPR/Cas9 system. We demonstrate that CRISPR/Cas9-mediated excision of a long inhibitory sequence from Gdnf native 3'UTR in mouse zygotes increases the levels of endogenous GDNF with similar phenotypic alterations in embryonic kidney development as we described in GDNF constitutive and conditional hypermorphic mice. Furthermore, we show that CRISPR/Cas9-mediated targeting of 3’UTRs in vivo allows the modulation of the expression levels of two other morphogens, Gdf11 and Bdnf. Together, our work demonstrates the power of in vivo 3’UTR editing using the CRISPR/Cas9 system to create hyper- and hypomorphic alleles, suggesting wide applicability in studies on gene function and potentially, in gene therapy. Elsevier 2023-02-24 /pmc/articles/PMC10009458/ /pubmed/36923835 http://dx.doi.org/10.1016/j.heliyon.2023.e13844 Text en © 2023 Published by Elsevier Ltd. 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 Research Article
Mätlik, Kärt
Olfat, Soophie
Cowlishaw, Mark Cary
Moreno, Eva Domenech
Ollila, Saara
Andressoo, Jaan-Olle
In vivo modulation of endogenous gene expression via CRISPR/Cas9-mediated 3’UTR editing
title In vivo modulation of endogenous gene expression via CRISPR/Cas9-mediated 3’UTR editing
title_full In vivo modulation of endogenous gene expression via CRISPR/Cas9-mediated 3’UTR editing
title_fullStr In vivo modulation of endogenous gene expression via CRISPR/Cas9-mediated 3’UTR editing
title_full_unstemmed In vivo modulation of endogenous gene expression via CRISPR/Cas9-mediated 3’UTR editing
title_short In vivo modulation of endogenous gene expression via CRISPR/Cas9-mediated 3’UTR editing
title_sort in vivo modulation of endogenous gene expression via crispr/cas9-mediated 3’utr editing
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10009458/
https://www.ncbi.nlm.nih.gov/pubmed/36923835
http://dx.doi.org/10.1016/j.heliyon.2023.e13844
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