{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T22:32:51Z","timestamp":1786746771581,"version":"build-2736575974"},"posted":{"date-parts":[[2026,8,11]]},"group-title":"Developmental Biology","reference-count":55,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T00:00:00Z","timestamp":1786406400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"id":[{"id":"https:\/\/ror.org\/052zd4v68","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"crossref","award":["R35GM158421"],"award-info":[{"award-number":["R35GM158421"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Eunice Kennedy Shriver National Institute of Child Health and Human Development","award":["R01HD104825"],"award-info":[{"award-number":["R01HD104825"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,8,11]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  The tunicate\n                  <jats:italic>Ciona robusta<\/jats:italic>\n                  offers a tractable non-vertebrate chordate model for probing gene function via tissue-specific, CRISPR\/Cas9-mediated mutagenesis in F0. Building on Arcadia Science\u2019s Zoogle platform, which identifies and ranks orthologs of human genes from various non-traditional model organisms, we carried out a pilot project to probe the developmental roles of three notochord- and endoderm-expressed candidate orthologs of human disease genes (\n                  <jats:italic>Fcho<\/jats:italic>\n                  ,\n                  <jats:italic>Pgm3<\/jats:italic>\n                  , and\n                  <jats:italic>Nckap1<\/jats:italic>\n                  ) alongside a fourth gene (\n                  <jats:italic>Plastin<\/jats:italic>\n                  ) implicated in papilla cell elongation. This preprint compiles and updates a series of research project milestones previously posted episodically on Zenodo. Here we summarize the full results and our conclusion about this pilot project. Using CRISPR\/Cas9, we found that tissue-specific knockout of\n                  <jats:italic>Pgm3<\/jats:italic>\n                  and, to a lesser extent,\n                  <jats:italic>Fcho<\/jats:italic>\n                  caused significant defects in larval tail elongation. Separately, CRISPR knockout of\n                  <jats:italic>Plastin<\/jats:italic>\n                  , an actin-bundling gene expressed throughout the sensory-adhesive papillae of the larva, caused a subtle reduction in papilla cell elongation when combined as a duoble knockout with another actin-bundling protein-encoding gene,\n                  <jats:italic>Villin<\/jats:italic>\n                  . These results identify\n                  <jats:italic>Pgm3<\/jats:italic>\n                  as the most promising candidate for further development as a Ciona-based model of human disease and demonstrate the utility of tissue-specific CRISPR screening for prioritizing candidate disease gene orthologs identified through comparative genomics platforms like Zoogle.\n                <\/jats:p>","DOI":"10.64898\/2026.08.10.743552","type":"posted-content","created":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T18:05:26Z","timestamp":1786471526000},"source":"Crossref","is-referenced-by-count":0,"title":["Using CRISPR\/Cas9 to investigate the role of candidate human disease gene orthologs in Ciona"],"prefix":"10.64898","author":[{"given":"Sabrina A.","family":"Hernandez","sequence":"first","affiliation":[{"name":"Georgia Institute of Technology, School of Biological Sciences"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christopher J.","family":"Johnson","sequence":"additional","affiliation":[{"name":"Georgia Institute of Technology, School of Biological Sciences"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7179-9700","authenticated-orcid":false,"given":"Alberto","family":"Stolfi","sequence":"additional","affiliation":[{"name":"Georgia Institute of Technology, School of Biological Sciences"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026081414550848000_2026.08.10.743552v1.1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-cellbio-111315-125218"},{"key":"2026081414550848000_2026.08.10.743552v1.2","doi-asserted-by":"crossref","first-page":"2317","DOI":"10.1016\/j.jaci.2019.02.014","article-title":"F-BAR domain only protein 1 (FCHO1) deficiency is a novel cause of combined immune deficiency in human subjects","volume":"143","year":"2019","journal-title":"Journal of Allergy and Clinical Immunology"},{"key":"2026081414550848000_2026.08.10.743552v1.3","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-019-1385-y"},{"key":"2026081414550848000_2026.08.10.743552v1.4","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20192275"},{"key":"2026081414550848000_2026.08.10.743552v1.5","unstructured":"Chou, S. , Patton, A.H. , Sun, D.A ., 2025. 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