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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...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
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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. |
format | Online Article Text |
id | pubmed-6523736 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
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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