CRISPR/Cas9-mediated knock-out of dUTPase in mice leads to early embryonic lethality

• DOI: 10.1101/335422
• Publication Date: 2018-05-31T13:45:30Z
• Authors: Hajnalka Laura Pá...; Gergely Rácz; Zoltán Gál; Orsolya Hoffmann; Gergely Tihanyi; Elen Gócza; László Hiripi; Beáta G. Vértessy
• Keywords: Synthetic Lethality; Gene knockin

Abstract Sanitization of nucleotide pools is essential for genome maintenance. Among the enzymes significant in this mechanism, deoxyuridine 5′-triphosphate nucleotidohydrolase (dUTPase) performs cleavage dUTP into dUMP and inorganic pyrophosphate. By reaction enzyme efficiently prevents uracil incorporation DNA provides dUMP, substrate de novo thymidylate biosynthesis. Despite its physiological significance, knock-out models dUTPase have not yet been investigated mammals, only unicellular organisms, such as bacteria yeast. Here we generate CRISPR/Cas9-mediated mice. We find that heterozygous dut +/-animals are viable while decreased level clearly observable. also show embryonic development. Based on present results, early -/-embryos can still reach blastocyst stage, however, they die shortly after implantation. Analysis preimplantion embryos indicate perturbed growth both inner cell mass (ICM) trophectoderm (TE). conclude indispensable post-implantation development The gene targeting model generated study will allow further detailed studies combination with additional knock-outs.