{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,20]],"date-time":"2026-04-20T14:49:26Z","timestamp":1776696566308,"version":"3.51.2"},"reference-count":27,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2021,4,15]],"date-time":"2021-04-15T00:00:00Z","timestamp":1618444800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2021,4,15]],"date-time":"2021-04-15T00:00:00Z","timestamp":1618444800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"name":"JSPS KAKENHI","award":["16K07091"],"award-info":[{"award-number":["16K07091"]}]},{"name":"Start Up Fund for female researchers in NAIST"},{"name":"KAC 40th Anniversary Research Grant"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    E26 avian leukemia oncogene 2, 3\u2032 domain (Ets2) has been implicated in various biological processes. An\n                    <jats:italic>Ets2<\/jats:italic>\n                    mutant model (\n                    <jats:italic>Ets2<\/jats:italic>\n                    <jats:sup>\n                      <jats:italic>db1\/db1<\/jats:italic>\n                    <\/jats:sup>\n                    ), which lacks the DNA-binding domain, was previously reported to exhibit embryonic lethality caused by a trophoblast abnormality. This phenotype could be rescued by tetraploid complementation, resulting in pups with wavy hair and curly whiskers. Here, we generated new\n                    <jats:italic>Ets2<\/jats:italic>\n                    mutant models with a frame-shift mutation in exon 8 using the CRISPR\/Cas9 method. Homozygous mutants could not be obtained by natural mating as embryonic development stopped before E8.5, as previously reported. When we rescued them by tetraploid complementation, these mice did not exhibit wavy hair or curly whisker phenotypes. Our newly generated mice exhibited exon 8 skipping, which led to in-frame mutant mRNA expression in the skin and thymus but not in E7.5\n                    <jats:italic>Ets2<\/jats:italic>\n                    <jats:sup>\n                      <jats:italic>em1\/em1<\/jats:italic>\n                    <\/jats:sup>\n                    embryos. This exon 8-skipped\n                    <jats:italic>Ets2<\/jats:italic>\n                    mRNA was translated into protein, suggesting that this Ets2 mutant protein complemented the Ets2 function in the skin. Our data implies that novel splicing variants incidentally generated after genome editing may complicate the phenotypic analysis but may also give insight into the new mechanisms related to biological gene functions.\n                  <\/jats:p>","DOI":"10.1038\/s41598-021-87751-5","type":"journal-article","created":{"date-parts":[[2021,4,15]],"date-time":"2021-04-15T06:18:05Z","timestamp":1618467485000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["A novel tissue specific alternative splicing variant mitigates phenotypes in\u00a0Ets2\u00a0frame-shift mutant models"],"prefix":"10.1038","volume":"11","author":[{"given":"Yuki","family":"Kishimoto","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Iori","family":"Nishiura","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wataru","family":"Hirata","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shunsuke","family":"Yuri","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nami","family":"Yamamoto","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Masahito","family":"Ikawa","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ayako","family":"Isotani","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2021,4,15]]},"reference":[{"key":"87751_CR1","doi-asserted-by":"publisher","first-page":"1451","DOI":"10.1101\/gad.4.9.1451","volume":"4","author":"FD Karim","year":"1990","unstructured":"Karim, F. D. et al. The ETS-domain: A new DNA-binding motif that recognizes a purine-rich core DNA sequence. Genes Dev. 4, 1451\u20131453. https:\/\/doi.org\/10.1101\/gad.4.9.1451 (1990).","journal-title":"Genes Dev."},{"key":"87751_CR2","doi-asserted-by":"publisher","first-page":"827","DOI":"10.1038\/35099076","volume":"2","author":"AD Sharrocks","year":"2001","unstructured":"Sharrocks, A. D. The ETS-domain transcription factor family. Nat. Rev. Mol. Cell Biol. 2, 827\u2013837. https:\/\/doi.org\/10.1038\/35099076 (2001).","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"87751_CR3","doi-asserted-by":"publisher","first-page":"127","DOI":"10.1101\/gad.950902","volume":"16","author":"JJ Seidel","year":"2002","unstructured":"Seidel, J. J. & Graves, B. J. An ERK2 docking site in the Pointed domain distinguishes a subset of ETS transcription factors. Genes Dev. 16, 127\u2013137. https:\/\/doi.org\/10.1101\/gad.950902 (2002).","journal-title":"Genes Dev."},{"key":"87751_CR4","doi-asserted-by":"publisher","first-page":"1315","DOI":"10.1101\/gad.12.9.1315","volume":"12","author":"H Yamamoto","year":"1998","unstructured":"Yamamoto, H. et al. Defective trophoblast function in mice with a targeted mutation of Ets2. Genes Dev. 12, 1315\u20131326. https:\/\/doi.org\/10.1101\/gad.12.9.1315 (1998).","journal-title":"Genes Dev."},{"key":"87751_CR5","doi-asserted-by":"publisher","first-page":"8614","DOI":"10.1128\/mcb.23.23.8614-8625.2003","volume":"23","author":"AK Man","year":"2003","unstructured":"Man, A. K. et al. Ets2-dependent stromal regulation of mouse mammary tumors. Mol. Cell Biol. 23, 8614\u20138625. https:\/\/doi.org\/10.1128\/mcb.23.23.8614-8625.2003 (2003).","journal-title":"Mol. Cell Biol."},{"key":"87751_CR6","doi-asserted-by":"publisher","first-page":"1123","DOI":"10.1182\/blood-2009-03-211391","volume":"114","author":"G Wei","year":"2009","unstructured":"Wei, G. et al. Ets1 and Ets2 are required for endothelial cell survival during embryonic angiogenesis. Blood 114, 1123\u20131130. https:\/\/doi.org\/10.1182\/blood-2009-03-211391 (2009).","journal-title":"Blood"},{"key":"87751_CR7","doi-asserted-by":"publisher","first-page":"999","DOI":"10.1165\/rcmb.2010-0490OC","volume":"45","author":"CP Baran","year":"2011","unstructured":"Baran, C. P. et al. Transcription factor ets-2 plays an important role in the pathogenesis of pulmonary fibrosis. Am. J. Respir. Cell Mol. Biol. 45, 999\u20131006. https:\/\/doi.org\/10.1165\/rcmb.2010-0490OC (2011).","journal-title":"Am. J. Respir. Cell Mol. 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Multiplex genome engineering using CRISPR\/Cas systems. Science 339, 819\u2013823. https:\/\/doi.org\/10.1126\/science.1231143 (2013).","journal-title":"Science"},{"key":"87751_CR11","doi-asserted-by":"publisher","first-page":"823","DOI":"10.1126\/science.1232033","volume":"339","author":"P Mali","year":"2013","unstructured":"Mali, P. et al. RNA-guided human genome engineering via Cas9. Science 339, 823\u2013826. https:\/\/doi.org\/10.1126\/science.1232033 (2013).","journal-title":"Science"},{"key":"87751_CR12","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.ydbio.2016.07.017","volume":"418","author":"M Hashimoto","year":"2016","unstructured":"Hashimoto, M., Yamashita, Y. & Takemoto, T. Electroporation of Cas9 protein\/sgRNA into early pronuclear zygotes generates non-mosaic mutants in the mouse. Dev. Biol. 418, 1\u20139. https:\/\/doi.org\/10.1016\/j.ydbio.2016.07.017 (2016).","journal-title":"Dev. 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Genet."},{"key":"87751_CR16","doi-asserted-by":"publisher","first-page":"437","DOI":"10.1038\/nrm.2017.27","volume":"18","author":"FE Baralle","year":"2017","unstructured":"Baralle, F. E. & Giudice, J. Alternative splicing as a regulator of development and tissue identity. Nat. Rev. Mol. Cell Biol. 18, 437\u2013451. https:\/\/doi.org\/10.1038\/nrm.2017.27 (2017).","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"87751_CR17","doi-asserted-by":"publisher","first-page":"108","DOI":"10.1186\/s13059-017-1237-8","volume":"18","author":"H Mou","year":"2017","unstructured":"Mou, H. et al. CRISPR\/Cas9-mediated genome editing induces exon skipping by alternative splicing or exon deletion. Genome Biol. 18, 108. https:\/\/doi.org\/10.1186\/s13059-017-1237-8 (2017).","journal-title":"Genome Biol."},{"key":"87751_CR18","doi-asserted-by":"publisher","first-page":"60","DOI":"10.1186\/s12896-018-0465-7","volume":"18","author":"D Chen","year":"2018","unstructured":"Chen, D. et al. CRISPR\/Cas9-mediated genome editing induces exon skipping by complete or stochastic altering splicing in the migratory locust. BMC Biotechnol. 18, 60. https:\/\/doi.org\/10.1186\/s12896-018-0465-7 (2018).","journal-title":"BMC Biotechnol."},{"key":"87751_CR19","doi-asserted-by":"publisher","first-page":"164","DOI":"10.1186\/s13059-018-1532-z","volume":"19","author":"T Sui","year":"2018","unstructured":"Sui, T. et al. CRISPR-induced exon skipping is dependent on premature termination codon mutations. Genome Biol. 19, 164. https:\/\/doi.org\/10.1186\/s13059-018-1532-z (2018).","journal-title":"Genome Biol."},{"key":"87751_CR20","doi-asserted-by":"publisher","first-page":"4056","DOI":"10.1038\/s41467-019-12028-5","volume":"10","author":"R Tuladhar","year":"2019","unstructured":"Tuladhar, R. et al. CRISPR-Cas9-based mutagenesis frequently provokes on-target mRNA misregulation. Nat. Commun. 10, 4056. https:\/\/doi.org\/10.1038\/s41467-019-12028-5 (2019).","journal-title":"Nat. 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Nature 548, 219\u2013223. https:\/\/doi.org\/10.1038\/nature23274 (2017).","journal-title":"Nature"},{"key":"87751_CR25","doi-asserted-by":"publisher","first-page":"31666","DOI":"10.1038\/srep31666","volume":"6","author":"A Oji","year":"2016","unstructured":"Oji, A. et al. CRISPR\/Cas9 mediated genome editing in ES cells and its application for chimeric analysis in mice. Sci. Rep. 6, 31666. https:\/\/doi.org\/10.1038\/srep31666 (2016).","journal-title":"Sci. Rep."},{"key":"87751_CR26","doi-asserted-by":"publisher","first-page":"946","DOI":"10.1038\/ng.397","volume":"41","author":"C Kokubu","year":"2009","unstructured":"Kokubu, C. et al. A transposon-based chromosomal engineering method to survey a large cis-regulatory landscape in mice. Nat. Genet. 41, 946\u2013952. https:\/\/doi.org\/10.1038\/ng.397 (2009).","journal-title":"Nat. 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