{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T00:41:50Z","timestamp":1787013710453,"version":"build-2736575974"},"reference-count":41,"publisher":"Wiley","license":[{"start":{"date-parts":[[2025,10,28]],"date-time":"2025-10-28T00:00:00Z","timestamp":1761609600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Insect Science"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Knock\u2010in efficiency\n                    <jats:italic>via<\/jats:italic>\n                    CRISPR\/Cas9\u2010based homology\u2010directed repair (HDR) is low in lepidopteran insects due to predominance of knockout through non\u2010homologous end joining (NHEJ) pathway. DNA ligase 4 (LIG4), a core protein of NHEJ complex and double\u2010strand break (DSB) binding repair, is a promising target for enhancing HDR efficiency in diverse species. However, the role of Lig4 in modulating HDR efficiency in\n                    <jats:italic>Plutella xylostella<\/jats:italic>\n                    , a highly destructive pest of vegetable crops, remains unclear. The\n                    <jats:italic>PxLig4<\/jats:italic>\n                    gene was identified from the\n                    <jats:italic>P. xylostella<\/jats:italic>\n                    genome with 4 conserved domains: ATP\u2010dependent DNA ligase core\u2010adenylation middle domain, ATP\u2010dependent DNA ligase core\u2011adenylation N\u2010terminal domain, ATP\u2010dependent DNA ligase core\u2011adenylation C\u2011terminal domain, and BRCA1 C terminus domain. Multiple sequence alignment and phylogenetic analysis confirmed strong conservation of PxLig4 among Lepidoptera. Real\u2010time quantitative polymerase chain reaction (RT\u2010qPCR) analysis showed that\n                    <jats:italic>PxLig4<\/jats:italic>\n                    was highly expressed in eggs, female adults, and male adults, particularly in the ovary, followed by the testis, fat body, and midgut tissues. A homozygous\n                    <jats:italic>PxLig4<\/jats:italic>\n                    knockout line (\u0394\n                    <jats:italic>PxLig4<\/jats:italic>\n                    ) with a 2\u2010bp deletion was successfully generated by using the CRISPR\/Cas9 technique. \u0394\n                    <jats:italic>PxLig4<\/jats:italic>\n                    exhibited significant developmental and reproductive defects. A donor vector to knock in the enhanced green fluorescent protein (\n                    <jats:italic>EGFP<\/jats:italic>\n                    ) gene at the eye color gene\n                    <jats:italic>PxKmo<\/jats:italic>\n                    was developed to assess knock\u2010in efficiency. The knock\u2010in efficiency in \u0394\n                    <jats:italic>PxLig4<\/jats:italic>\n                    was 2.86%, significantly higher than the 0.26% observed in the wild\u2010type\n                    <jats:italic>P. xylostella<\/jats:italic>\n                    . These results implicate Lig4 as a supporting factor for NHEJ and demonstrate that suppression of\n                    <jats:italic>PxLig4<\/jats:italic>\n                    enhances HDR efficiency in\n                    <jats:italic>P. xylostella<\/jats:italic>\n                    , providing a potential strategy to improve the efficiency of gene knock\u2010in in insect pests.\n                  <\/jats:p>","DOI":"10.1111\/1744-7917.70184","type":"journal-article","created":{"date-parts":[[2025,10,28]],"date-time":"2025-10-28T13:33:59Z","timestamp":1761658439000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Knockout of\n                    <i>Ligase4<\/i>\n                    gene enhances homology\u2010directed repair efficiency in\n                    <i>Plutella xylostella<\/i>"],"prefix":"10.1111","author":[{"ORCID":"https:\/\/orcid.org\/0009-0002-0053-8452","authenticated-orcid":false,"given":"Faisal","family":"Munir","sequence":"first","affiliation":[{"name":"State Key Laboratory of Agricultural and Forestry Biosecurity, Institute of Applied Ecology Fujian Agriculture and Forestry University  Fuzhou China"},{"name":"Joint International Research Laboratory of Ecological Pest Control Ministry of Education  Fuzhou China"},{"name":"Ministerial and Provincial Joint Innovation Centre for Safety Production of Cross\u2010Strait Crops Fujian Agriculture and Forestry University  Fuzhou China"},{"name":"Key Laboratory of Integrated Pest Management for Fujian\u2010Taiwan Crops Ministry of Agriculture  Fuzhou China"},{"name":"Key Laboratory of Green Pest Control Fujian Province University  Fuzhou China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Uroosa","family":"Zaheer","sequence":"additional","affiliation":[{"name":"State Key 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