{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T08:07:50Z","timestamp":1789027670237,"version":"build-2803163510"},"reference-count":33,"publisher":"Cambridge University Press (CUP)","license":[{"start":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T00:00:00Z","timestamp":1788998400000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/www.cambridge.org\/core\/terms"}],"funder":[{"DOI":"10.13039\/501100001503","name":"Indian Council of Agricultural Research","doi-asserted-by":"publisher","award":["ICAR CABin"],"award-info":[{"award-number":["ICAR CABin"]}],"id":[{"id":"10.13039\/501100001503","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["cambridge.org"],"crossmark-restriction":true},"short-container-title":["Bull. Entomol. Res."],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    <jats:italic>Maruca vitrata<\/jats:italic>\n                    , the legume pod borer, causes yield losses of up to 80% in grain legumes. Increasing insecticide resistance and environmental concerns necessitate sustainable pest management alternatives. In the present study, the complete vitellogenin (Vg) coding sequence (CDS), a key reproductive gene involved in oogenesis and embryonic development, was cloned and molecularly characterised from\n                    <jats:italic>M. vitrata<\/jats:italic>\n                    . The assembled Vg CDS (\u223c5.3 kb) shared 99.04% sequence identity with the reported\n                    <jats:italic>M. vitrata<\/jats:italic>\n                    Vg sequence (MG799570.1). Phylogenetic analysis demonstrated close evolutionary association with related Lepidopteran species, while protein domain analysis identified three conserved domains, namely LPD_N, DUF1943, and VWD. Among these, the single exon-encoded LPD_N domain was selected as the target region for CRISPR\/Cas9-mediated editing. Homology models of Vg and vitellogenin receptor (VgR) (Global Model Quality Estimation (GMQE): 0.58 and 0.51) showed a favourable interaction by protein\u2013protein docking (score: \u2212295.66). Three single-guide RNAs (sgRNAs) were designed, synthesised through in-vitro transcription, and evaluated using in vitro cleavage assays. sgRNA1 targeting the LPD_N domain and sgRNA2 targeting the signal peptide region exhibited efficient site-specific cleavage activity, whereas sgRNA3 failed to induce cleavage because of an unfavourable secondary structure that likely impaired Cas9-sgRNA complex formation. Overall, this study provides the first CRISPR-oriented functional characterisation and sgRNA validation of the\n                    <jats:italic>M. vitrata<\/jats:italic>\n                    Vg gene, together with structural characterisation of VgR and Vg-VgR interaction analysis, providing preliminary molecular resources for future CRISPR\/Cas9 studies and supporting future embryo microinjection and heritable\u00a0genome editing for sustainable management of\n                    <jats:italic>M. vitrata<\/jats:italic>\n                    .\n                  <\/jats:p>","DOI":"10.1017\/s0007485326101308","type":"journal-article","created":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T07:32:28Z","timestamp":1789025548000},"page":"1-13","update-policy":"https:\/\/doi.org\/10.1017\/policypage","source":"Crossref","is-referenced-by-count":0,"title":["Molecular characterization of vitellogenin and its receptor with CRISPR-based sgRNA validation in the legume pod borer,\n                    <i>Maruca vitrata<\/i>\n                    (Geyer) (Lepidoptera: Crambidae)"],"prefix":"10.1017","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8402-424X","authenticated-orcid":false,"given":"Chalapathi","family":"Pradeep","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04ghh8334","id-type":"ROR","asserted-by":"publisher"}],"name":"Central Sericultural Research and Training Institute"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Parvathy Madhusoodanan","family":"Sujatha","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03js6zg56","id-type":"ROR","asserted-by":"publisher"}],"name":"University of Agricultural Sciences"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Somakalahalli Narasimhappa","family":"Nagesha","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/03js6zg56","id-type":"ROR","asserted-by":"publisher"}],"name":"University of Agricultural Sciences"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ramasamy","family":"Asokan","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00s2dqx11","id-type":"ROR","asserted-by":"publisher"}],"name":"ICAR-Indian Institute of Horticultural Research"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4094-6285","authenticated-orcid":false,"given":"Ashok","family":"Karuppannasamy","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/01bzgdw81","id-type":"ROR","asserted-by":"publisher"}],"name":"ICAR-Indian Agricultural Research Institute"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"56","published-online":{"date-parts":[[2026,9,10]]},"reference":[{"key":"S0007485326101308_ref26","doi-asserted-by":"publisher","DOI":"10.1093\/molbev\/msab120"},{"key":"S0007485326101308_ref3","doi-asserted-by":"publisher","DOI":"10.1016\/j.gene.2024.148925"},{"key":"S0007485326101308_ref12","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.2501"},{"key":"S0007485326101308_ref15","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-07964-7"},{"key":"S0007485326101308_ref32","doi-asserted-by":"publisher","DOI":"10.1002\/prot.25234"},{"key":"S0007485326101308_ref33","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-025-59035-3"},{"key":"S0007485326101308_ref24","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-ento-021220-084539"},{"key":"S0007485326101308_ref6","doi-asserted-by":"publisher","DOI":"10.1007\/s12298-022-01133-3"},{"key":"S0007485326101308_ref25","doi-asserted-by":"publisher","DOI":"10.1093\/bioinformatics\/btz828"},{"key":"S0007485326101308_ref5","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2015.04.028"},{"key":"S0007485326101308_ref9","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijbiomac.2025.146574"},{"key":"S0007485326101308_ref30","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkm290"},{"key":"S0007485326101308_ref17","doi-asserted-by":"crossref","first-page":"103500","DOI":"10.1016\/j.dci.2019.103500","article-title":"Deletion mutant of sPLA2 using CRISPR\/Cas9 exhibits immunosuppression, developmental retardation, and failure of oocyte development in legume pod borer, Maruca vitrata","volume":"103","author":"Lee","year":"2020","journal-title":"Developmental and Comparative Immunology"},{"key":"S0007485326101308_ref28","doi-asserted-by":"publisher","DOI":"10.1007\/s10340-025-01939-6"},{"key":"S0007485326101308_ref8","first-page":"82","article-title":"Pymol: An open-source molecular graphics tool","volume":"40","author":"DeLano","year":"2002","journal-title":"CCP4 Newsl. 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