{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T23:19:41Z","timestamp":1768519181336,"version":"3.49.0"},"posted":{"date-parts":[[2024,11,15]]},"group-title":"Genetics","reference-count":48,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2024,11,15]],"date-time":"2024-11-15T00:00:00Z","timestamp":1731628800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2024,11,19]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Peroxisomal biogenesis disorders (PBD) are autosomal recessive disorders caused by loss-of-function mutations of one of the\n                  <jats:italic>PEX<\/jats:italic>\n                  genes responsible for peroxisomal formation. Impaired peroxisome assembly causes severe multisystemic failure with patient phenotypes ranging from epilepsy, liver disease, feeding issues, biochemical abnormalities, and neurodegeneration. Variants in the same\n                  <jats:italic>PEX<\/jats:italic>\n                  gene can produce wide differences in severity, ranging from individuals with death in the first year of life to adults with milder complications. To study this strong genotype-phenotype correlation, we selected specific human\n                  <jats:italic>PEX<\/jats:italic>\n                  gene mutations and utilized\n                  <jats:italic>Drosophila<\/jats:italic>\n                  as a model organism. We generated flies replacing the coding sequence of our\n                  <jats:italic>Pex<\/jats:italic>\n                  gene of interest with a\n                  <jats:italic>KozakGAL4 (KZ)<\/jats:italic>\n                  promoter trap sequence. These cassettes simultaneously knock-out of the\n                  <jats:italic>Pex<\/jats:italic>\n                  gene and knock-in a\n                  <jats:italic>GAL4<\/jats:italic>\n                  driver, ideal for making \u201chumanized\u201d flies in which the human\n                  <jats:italic>PEX<\/jats:italic>\n                  gene can replace the fly loss. We assessed\n                  <jats:italic>\n                    Pex2\n                    <jats:sup>KZ<\/jats:sup>\n                  <\/jats:italic>\n                  and\n                  <jats:italic>\n                    Pex16\n                    <jats:sup>KZ<\/jats:sup>\n                  <\/jats:italic>\n                  lines in lifespan, bang sensitivity, and climbing assays and confirmed that these are strong loss-of-function alleles. In parallel, we generated human reference and variant UAS-cDNA lines of\n                  <jats:italic>PEX2<\/jats:italic>\n                  and\n                  <jats:italic>PEX16<\/jats:italic>\n                  variants in\n                  <jats:italic>Drosophila<\/jats:italic>\n                  . We observed nearly complete phenotypic rescue of\n                  <jats:italic>Drosophila Pex2<\/jats:italic>\n                  and\n                  <jats:italic>Pex16<\/jats:italic>\n                  loss when human\n                  <jats:italic>\n                    PEX2\n                    <jats:sup>Ref<\/jats:sup>\n                  <\/jats:italic>\n                  or\n                  <jats:italic>\n                    PEX16\n                    <jats:sup>Ref<\/jats:sup>\n                  <\/jats:italic>\n                  , respectively, were expressed. We also provide evidence for an allele severity spectrum in\n                  <jats:italic>PEX2<\/jats:italic>\n                  and\n                  <jats:italic>PEX16<\/jats:italic>\n                  in which some missense alleles, such as\n                  <jats:italic>\n                    PEX2\n                    <jats:sup>C247R<\/jats:sup>\n                  <\/jats:italic>\n                  , are equally severe as early truncations, such as\n                  <jats:italic>\n                    PEX2\n                    <jats:sup>R119*<\/jats:sup>\n                  <\/jats:italic>\n                  . We also observed that alleles associated with mild PBD, such as\n                  <jats:italic>\n                    PEX2\n                    <jats:sup>E55K<\/jats:sup>\n                  <\/jats:italic>\n                  , show variability depending on the assay but do not fully rescue. Finally, alleles associated with atypical ataxia phenotypes, such as\n                  <jats:italic>\n                    PEX16\n                    <jats:sup>F332Del<\/jats:sup>\n                  <\/jats:italic>\n                  , can perform as well as\n                  <jats:italic>\n                    PEX16\n                    <jats:sup>Ref<\/jats:sup>\n                  <\/jats:italic>\n                  , depending on the assay. Altogether, these\n                  <jats:italic>Drosophila<\/jats:italic>\n                  lines effectively model the range of severity of peroxisomal biogenesis disorders.\n                <\/jats:p>","DOI":"10.1101\/2024.11.14.623590","type":"posted-content","created":{"date-parts":[[2024,11,15]],"date-time":"2024-11-15T23:15:25Z","timestamp":1731712525000},"source":"Crossref","is-referenced-by-count":0,"title":["Distinguishing\n                  <i>PEX<\/i>\n                  gene variant severity for mild, severe, and atypical peroxisome biogenesis disorders in\n                  <i>Drosophila<\/i>"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-0617-9127","authenticated-orcid":false,"given":"Vanessa A.","family":"Gomez","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Oguz","family":"Kanca","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7389-0890","authenticated-orcid":false,"given":"Sharayu V.","family":"Jangam","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Saurabh","family":"Srivastav","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3086-7225","authenticated-orcid":false,"given":"Jonathan C.","family":"Andrews","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5245-5910","authenticated-orcid":false,"given":"Michael F.","family":"Wangler","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2024112109150674000_2024.11.14.623590v2.1","doi-asserted-by":"publisher","DOI":"10.59566\/IJBS.2006.2308"},{"key":"2024112109150674000_2024.11.14.623590v2.2","doi-asserted-by":"crossref","unstructured":"Wanders, R. 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