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CRISPR/Cas9-Mediated Knock-Out of dUTPase in Mice Leads to Early Embryonic Lethality

Sanitization of nucleotide pools is essential for genome maintenance. Deoxyuridine 5′-triphosphate nucleotidohydrolase (dUTPase) is a key enzyme in this pathway since it catalyzes the cleavage of 2′-deoxyuridine 5′-triphosphate (dUTP) into 2′-deoxyuridine 5′-monophosphate (dUMP) and inorganic pyroph...

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Autores principales: Pálinkás, Hajnalka Laura, Rácz, Gergely Attila, Gál, Zoltán, Hoffmann, Orsolya Ivett, Tihanyi, Gergely, Róna, Gergely, Gócza, Elen, Hiripi, László, Vértessy, Beáta G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6523736/
https://www.ncbi.nlm.nih.gov/pubmed/30987342
http://dx.doi.org/10.3390/biom9040136
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author Pálinkás, Hajnalka Laura
Rácz, Gergely Attila
Gál, Zoltán
Hoffmann, Orsolya Ivett
Tihanyi, Gergely
Róna, Gergely
Gócza, Elen
Hiripi, László
Vértessy, Beáta G.
author_facet Pálinkás, Hajnalka Laura
Rácz, Gergely Attila
Gál, Zoltán
Hoffmann, Orsolya Ivett
Tihanyi, Gergely
Róna, Gergely
Gócza, Elen
Hiripi, László
Vértessy, Beáta G.
author_sort Pálinkás, Hajnalka Laura
collection PubMed
description Sanitization of nucleotide pools is essential for genome maintenance. Deoxyuridine 5′-triphosphate nucleotidohydrolase (dUTPase) is a key enzyme in this pathway since it catalyzes the cleavage of 2′-deoxyuridine 5′-triphosphate (dUTP) into 2′-deoxyuridine 5′-monophosphate (dUMP) and inorganic pyrophosphate. Through its action dUTPase efficiently prevents uracil misincorporation into DNA and at the same time provides dUMP, the substrate for de novo thymidylate biosynthesis. Despite its physiological significance, knock-out models of dUTPase have not yet been investigated in mammals, but only in unicellular organisms, such as bacteria and yeast. Here we generate CRISPR/Cas9-mediated dUTPase knock-out in mice. We find that heterozygous dut +/– animals are viable while having decreased dUTPase levels. Importantly, we show that dUTPase is essential for embryonic development since early dut −/− embryos reach the blastocyst stage, however, they die shortly after implantation. Analysis of pre-implantation embryos indicates perturbed growth of both inner cell mass (ICM) and trophectoderm (TE). We conclude that dUTPase is indispensable for post-implantation development in mice.
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spelling pubmed-65237362019-06-03 CRISPR/Cas9-Mediated Knock-Out of dUTPase in Mice Leads to Early Embryonic Lethality Pálinkás, Hajnalka Laura Rácz, Gergely Attila Gál, Zoltán Hoffmann, Orsolya Ivett Tihanyi, Gergely Róna, Gergely Gócza, Elen Hiripi, László Vértessy, Beáta G. Biomolecules Article Sanitization of nucleotide pools is essential for genome maintenance. Deoxyuridine 5′-triphosphate nucleotidohydrolase (dUTPase) is a key enzyme in this pathway since it catalyzes the cleavage of 2′-deoxyuridine 5′-triphosphate (dUTP) into 2′-deoxyuridine 5′-monophosphate (dUMP) and inorganic pyrophosphate. Through its action dUTPase efficiently prevents uracil misincorporation into DNA and at the same time provides dUMP, the substrate for de novo thymidylate biosynthesis. Despite its physiological significance, knock-out models of dUTPase have not yet been investigated in mammals, but only in unicellular organisms, such as bacteria and yeast. Here we generate CRISPR/Cas9-mediated dUTPase knock-out in mice. We find that heterozygous dut +/– animals are viable while having decreased dUTPase levels. Importantly, we show that dUTPase is essential for embryonic development since early dut −/− embryos reach the blastocyst stage, however, they die shortly after implantation. Analysis of pre-implantation embryos indicates perturbed growth of both inner cell mass (ICM) and trophectoderm (TE). We conclude that dUTPase is indispensable for post-implantation development in mice. MDPI 2019-04-04 /pmc/articles/PMC6523736/ /pubmed/30987342 http://dx.doi.org/10.3390/biom9040136 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Pálinkás, Hajnalka Laura
Rácz, Gergely Attila
Gál, Zoltán
Hoffmann, Orsolya Ivett
Tihanyi, Gergely
Róna, Gergely
Gócza, Elen
Hiripi, László
Vértessy, Beáta G.
CRISPR/Cas9-Mediated Knock-Out of dUTPase in Mice Leads to Early Embryonic Lethality
title CRISPR/Cas9-Mediated Knock-Out of dUTPase in Mice Leads to Early Embryonic Lethality
title_full CRISPR/Cas9-Mediated Knock-Out of dUTPase in Mice Leads to Early Embryonic Lethality
title_fullStr CRISPR/Cas9-Mediated Knock-Out of dUTPase in Mice Leads to Early Embryonic Lethality
title_full_unstemmed CRISPR/Cas9-Mediated Knock-Out of dUTPase in Mice Leads to Early Embryonic Lethality
title_short CRISPR/Cas9-Mediated Knock-Out of dUTPase in Mice Leads to Early Embryonic Lethality
title_sort crispr/cas9-mediated knock-out of dutpase in mice leads to early embryonic lethality
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6523736/
https://www.ncbi.nlm.nih.gov/pubmed/30987342
http://dx.doi.org/10.3390/biom9040136
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