{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,10]],"date-time":"2026-07-10T18:12:52Z","timestamp":1783707172596,"version":"3.55.0"},"posted":{"date-parts":[[2026,7,8]]},"group-title":"Developmental Biology","reference-count":51,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,7,8]],"date-time":"2026-07-08T00:00:00Z","timestamp":1783468800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,7,8]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Robust execution of developmental cell fates requires precise spatiotemporal control of the fate-defining regulators. In\n                  <jats:italic>Caenorhabditis elegans<\/jats:italic>\n                  , temporal patterning of larval hypodermal fates is governed by the heterochronic gene regulatory network, in which microRNAs act as major post-transcriptional regulators by silencing temporal transcripts through 3\n                  <jats:bold>\u2032<\/jats:bold>\n                  UTR-dependent repression. Here, we investigate\n                  <jats:italic>lin-66<\/jats:italic>\n                  , which encodes a nematode-specific cold shock domain protein previously implicated in heterochronic regulation and reported to associate with the miRISC effector protein AIN-1. Using targeted domain mutations and genetic analysis, we show that LIN-66 activity in the hypodermal cell-fate patterning requires its cold shock domain. Loss of\n                  <jats:italic>lin-66<\/jats:italic>\n                  causes persistent expression of LIN-14 and LIN-28, two early temporal regulators in the hypodermal seam cells that are canonical microRNA targets. Analysis indicates that\n                  <jats:italic>lin-66<\/jats:italic>\n                  function in seam-cell fate patterning does not depend on the native 3\n                  <jats:bold>\u2032<\/jats:bold>\n                  UTR sequences of\n                  <jats:italic>lin-14<\/jats:italic>\n                  or\n                  <jats:italic>lin-28<\/jats:italic>\n                  , distinguishing its activity from canonical microRNA repression.. Consistent However, consistent with the a broad functional overlap between LIN-66 function and microRNA-mediated regulation, hypodermal\n                  <jats:italic>lin-66<\/jats:italic>\n                  loss-of-function phenotypes are strongly enhanced by mutations in\n                  <jats:italic>alg-1<\/jats:italic>\n                  and\n                  <jats:italic>ain-1\/2<\/jats:italic>\n                  , which encode components of the microRNA-induced silencing complex. Moreover, loss of\n                  <jats:italic>lin-66<\/jats:italic>\n                  enhances phenotypes in mutants sensitized for microRNA activity outside the hypodermis. Together, these findings identify LIN-66 as a cold shock domain-dependent post-transcriptional regulator that safeguards developmental timing by limiting persistence of early fate regulators through mechanisms that intersect with, but are partly separable from, canonical 3\n                  <jats:bold>\u2032<\/jats:bold>\n                  UTR-mediated microRNA repression.\n                <\/jats:p>","DOI":"10.64898\/2026.07.07.737059","type":"posted-content","created":{"date-parts":[[2026,7,8]],"date-time":"2026-07-08T14:15:11Z","timestamp":1783520111000},"source":"Crossref","is-referenced-by-count":0,"title":["Cold Shock Domain Protein LIN-66 cooperates with microRNA-pathway buffering to safeguard developmental timing"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-4291-8786","authenticated-orcid":false,"given":"Reyyan","family":"Bulut","sequence":"first","affiliation":[{"name":"Program in Molecular Medicine, University of Massachusetts Chan Medical School, Worcester, MA 01605, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5562-5345","authenticated-orcid":false,"given":"Victor","family":"Ambros","sequence":"additional","affiliation":[{"name":"Program in Molecular Medicine, University of Massachusetts Chan Medical School, Worcester, MA 01605, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026071010153911000_2026.07.07.737059v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.devcel.2005.07.009"},{"key":"2026071010153911000_2026.07.07.737059v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/S1534-5807(03)00127-8"},{"key":"2026071010153911000_2026.07.07.737059v1.3","doi-asserted-by":"publisher","DOI":"10.1101\/gad.1.4.398"},{"key":"2026071010153911000_2026.07.07.737059v1.4","doi-asserted-by":"publisher","DOI":"10.1101\/gad.5.10.1825"},{"key":"2026071010153911000_2026.07.07.737059v1.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2018.03.006"},{"key":"2026071010153911000_2026.07.07.737059v1.6","doi-asserted-by":"publisher","DOI":"10.26508\/lsa.202402673"},{"key":"2026071010153911000_2026.07.07.737059v1.7","doi-asserted-by":"publisher","DOI":"10.1101\/2024.02.20.581241"},{"key":"2026071010153911000_2026.07.07.737059v1.8","doi-asserted-by":"publisher","DOI":"10.7554\/elife.68336"},{"key":"2026071010153911000_2026.07.07.737059v1.9","doi-asserted-by":"publisher","DOI":"10.1016\/j.molmed.2008.07.001"},{"key":"2026071010153911000_2026.07.07.737059v1.10","doi-asserted-by":"publisher","DOI":"10.1002\/0471142727.mb2603s109"},{"key":"2026071010153911000_2026.07.07.737059v1.11","doi-asserted-by":"publisher","DOI":"10.1534\/genetics.118.301532"},{"issue":"8","key":"2026071010153911000_2026.07.07.737059v1.12","first-page":"2507","article-title":"Reversal of cell fate determination in Caenorhabditis elegans vulval development. 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