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However, functional insights into the roles of Hox genes in primitive ametabolous insects, which represent the critical transition from aquatic crustaceans to winged insects, have been limited. In this study, we identified complete protein\u2010coding sequences of 10 Hox genes in the Zygentoma\n                    <jats:italic>Thermobia domestica<\/jats:italic>\n                    , and applied clustered regularly interspaced short palindromic repeats (CRISPR) \/ CRISPR\u2010associated nuclease 9 (Cas 9)\u00a0mediated gene knockout (KO) to decipher their functions. We found that the roles of\n                    <jats:italic>pb<\/jats:italic>\n                    ,\n                    <jats:italic>Dfd<\/jats:italic>\n                    , and\n                    <jats:italic>Scr<\/jats:italic>\n                    are vital in specifying the appendages of the head in\n                    <jats:italic>T. domestica<\/jats:italic>\n                    , and these roles are relatively conserved in crustaceans and winged insects.\n                    <jats:italic>Antp<\/jats:italic>\n                    is essential for the development of the prothorax segment and the first pair of legs in\n                    <jats:italic>T. domestica<\/jats:italic>\n                    .\n                    <jats:italic>Ubx<\/jats:italic>\n                    and\n                    <jats:italic>abd\u2010A<\/jats:italic>\n                    fully repress appendage development in the abdomen of\n                    <jats:italic>T. domestica<\/jats:italic>\n                    , which implies a functional switch from crustaceans to insects. Additionally, the role of\n                    <jats:italic>ftz<\/jats:italic>\n                    in segmenting the abdomen of\n                    <jats:italic>T. domestica<\/jats:italic>\n                    suggests it has acquired new functions in primitive insects, beyond its traditional Hox\u2010like roles. Although KOs of\n                    <jats:italic>lab<\/jats:italic>\n                    ,\n                    <jats:italic>Hox3<\/jats:italic>\n                    , and\n                    <jats:italic>Abd\u2010B<\/jats:italic>\n                    did not result in obvious external phenotypic changes, they led to a significant decrease in hatching rates and substantial deviations in daily survival numbers compared to the negative control. These findings underscore the indispensable roles of all Hox genes during the embryonic development of\n                    <jats:italic>T. domestica<\/jats:italic>\n                    . Our study sheds new light on the functional evolution of Hox genes in ametabolous insects and enhances our understanding of the genetic underpinnings of insect development and diversification.\n                  <\/jats:p>","DOI":"10.1111\/1744-7917.13504","type":"journal-article","created":{"date-parts":[[2025,2,20]],"date-time":"2025-02-20T01:45:09Z","timestamp":1740015909000},"page":"131-146","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["The role of Hox genes in shaping embryonic external morphology of the primitive insect\n                    <i>Thermobia domestica<\/i>\n                    (Zygentoma)"],"prefix":"10.1111","volume":"33","author":[{"given":"Hu\u2010Na","family":"Lu","sequence":"first","affiliation":[{"name":"Guangdong Provincial Key Laboratory of Insect Development Biology and Applied Technology, Institute of Insect Science and Technology, School of Life Sciences South China Normal University  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