{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T23:14:11Z","timestamp":1768518851851,"version":"3.49.0"},"posted":{"date-parts":[[2025,1,10]]},"group-title":"Genetics","reference-count":46,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,1,10]],"date-time":"2025-01-10T00:00:00Z","timestamp":1736467200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,1,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Genomic studies of autism and other neurodevelopmental disorders have identified several relevant protein-coding and noncoding variants. One gene with an excess of protein-coding\n                  <jats:italic>de novo<\/jats:italic>\n                  variants is\n                  <jats:italic>EBF3<\/jats:italic>\n                  that also is the gene underlying the Hypotonia, Ataxia, and Delayed Development Syndrome (HADDS). In previous work, we have identified noncoding\n                  <jats:italic>de novo<\/jats:italic>\n                  variants in an enhancer of\n                  <jats:italic>EBF3<\/jats:italic>\n                  called hs737 and further showed that there was an enrichment of deletions of this enhancer in individuals with neurodevelopmental disorders. In this present study, we generated a novel mouse line that deletes the highly conserved, orthologous mouse region of hs737 within the Rr169617 regulatory region, and characterized the molecular and phenotypic aspects of this mouse model. This line contains a 1,160 bp deletion within Rr169617 and through heterozygous crosses we found a deviation from Mendelian expectation (p = 0.02) with a significant depletion of the deletion allele (p = 5.8 \u00d7 10\n                  <jats:sup>-4<\/jats:sup>\n                  ).\n                  <jats:italic>\n                    Rr169617\n                    <jats:sup>+\/-<\/jats:sup>\n                  <\/jats:italic>\n                  mice had a reduction of\n                  <jats:italic>Ebf3<\/jats:italic>\n                  expression by 10% and\n                  <jats:italic>\n                    Rr169617\n                    <jats:sup>-\/-<\/jats:sup>\n                  <\/jats:italic>\n                  mice had a reduction of\n                  <jats:italic>Ebf3<\/jats:italic>\n                  expression by 20%. Differential expression analyses in E12.5 forebrain, midbrain, and hindbrain in\n                  <jats:italic>\n                    Rr169617\n                    <jats:sup>+\/+<\/jats:sup>\n                  <\/jats:italic>\n                  versus\n                  <jats:italic>\n                    Rr169617\n                    <jats:sup>-\/-<\/jats:sup>\n                  <\/jats:italic>\n                  mice identified dysregulated genes including histone genes\n                  <jats:italic>(<\/jats:italic>\n                  i.e.,\n                  <jats:italic>Hist1h1e<\/jats:italic>\n                  ,\n                  <jats:italic>Hist1h2bk<\/jats:italic>\n                  ,\n                  <jats:italic>Hist1h3i<\/jats:italic>\n                  ,\n                  <jats:italic>Hist1h2ao)<\/jats:italic>\n                  and other brain development related genes (e.g.,\n                  <jats:italic>Chd5<\/jats:italic>\n                  ,\n                  <jats:italic>Ntng1<\/jats:italic>\n                  ).\n                  <jats:italic>A priori<\/jats:italic>\n                  phenotyping analysis (open field, hole board and light\/dark transition) identified sex-specific differences in behavioral traits when comparing\n                  <jats:italic>\n                    Rr169617\n                    <jats:sup>-\/-<\/jats:sup>\n                  <\/jats:italic>\n                  males versus females; whereby, males were observed to be less mobile, move slower, and spend more time in the dark. Furthermore, both sexes when homozygous for the enhancer deletion displayed body composition differences when compared to wild-type mice. Overall, we show that deletion within Rr169617 reduces the expression of\n                  <jats:italic>Ebf3<\/jats:italic>\n                  and results in phenotypic outcomes consistent with potential sex specific behavioral differences. This enhancer deletion line provides a valuable resource for others interested in noncoding regions in neurodevelopmental disorders and\/or those interested in the gene regulatory network downstream of\n                  <jats:italic>Ebf3<\/jats:italic>\n                  .\n                <\/jats:p>","DOI":"10.1101\/2025.01.09.631762","type":"posted-content","created":{"date-parts":[[2025,1,11]],"date-time":"2025-01-11T11:30:15Z","timestamp":1736595015000},"source":"Crossref","is-referenced-by-count":1,"title":["Generation and Characterization of a Knockout Mouse of an Enhancer of\n                  <i>EBF3<\/i>"],"prefix":"10.64898","author":[{"given":"Emily Cordova","family":"Hurtado","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Janine M.","family":"Wotton","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexander","family":"Gulka","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Crystal","family":"Burke","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jeffrey K.","family":"Ng","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ibrahim","family":"Bah","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Juana","family":"Manuel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hillary","family":"Heins","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stephen A.","family":"Murray","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David U.","family":"Gorkin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jacqueline K.","family":"White","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kevin A.","family":"Peterson","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8246-6477","authenticated-orcid":false,"given":"Tychele N.","family":"Turner","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2025011108300791000_2025.01.09.631762v1.1","doi-asserted-by":"crossref","unstructured":"Bai, D. , Yip, B.H.K. , Windham, G.C. , Sourander, A. , Francis, R. , Yoffe, R. , Glasson, E. , Mahjani, B. , Suominen, A. , Leonard, H. , et al. 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