{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T19:48:51Z","timestamp":1768506531744,"version":"3.49.0"},"posted":{"date-parts":[[2023,6,13]]},"group-title":"Synthetic Biology","reference-count":64,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,6,13]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Rapid spread of insecticide resistance among anopheline mosquitoes threatens malaria elimination efforts, necessitating development of alternative vector control technologies. Sterile Insect Technique (SIT) has been successfully implemented in multiple insect pests to suppress field populations by the release of large numbers of sterile males, yet it has proven difficult to adapt to\n                  <jats:italic>Anopheles<\/jats:italic>\n                  vectors. Here we outline adaptation of a CRISPR-based genetic sterilization system to selectively ablate male sperm cells in the malaria mosquito\n                  <jats:italic>Anopheles gambiae<\/jats:italic>\n                  . We achieve robust mosaic biallelic mutagenesis of z\n                  <jats:italic>ero population growth<\/jats:italic>\n                  (\n                  <jats:italic>zpg<\/jats:italic>\n                  , a gene essential for differentiation of germ cells) in F1 individuals after intercrossing a germline-expressing Cas9 transgenic line to a line expressing\n                  <jats:italic>zpg<\/jats:italic>\n                  -targeting gRNAs. Approximately 95% of mutagenized males display complete genetic sterilization, and cause similarly high levels of infertility in their female mates. Using a fluorescence reporter that allows detection of the germline leads to a 100% accurate selection of spermless males, improving the system. These males cause a striking reduction in mosquito population size when released at field-like frequencies in competition cages against wild type males. These findings demonstrate that such a genetic system could be adopted for SIT against important malaria vectors.\n                <\/jats:p>","DOI":"10.1101\/2023.06.13.544841","type":"posted-content","created":{"date-parts":[[2023,6,14]],"date-time":"2023-06-14T00:15:13Z","timestamp":1686701713000},"source":"Crossref","is-referenced-by-count":4,"title":["CRISPR-mediated germline mutagenesis for genetic sterilization of\n                  <i>Anopheles gambiae<\/i>\n                  males"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3281-1161","authenticated-orcid":false,"given":"Andrea L.","family":"Smidler","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Douglas G.","family":"Paton","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"George M.","family":"Church","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kevin M.","family":"Esvelt","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2806-9718","authenticated-orcid":false,"given":"W. Robert","family":"Shaw","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Flaminia","family":"Catteruccia","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2023061611200760000_2023.06.13.544841v1.1","unstructured":"Organization WH. World malaria report 2021. 2021. Reference Source. 2022."},{"key":"2023061611200760000_2023.06.13.544841v1.2","unstructured":"Organization WH. World Malaria Report 2016 (World Health Organization, Geneva). 2016."},{"key":"2023061611200760000_2023.06.13.544841v1.3","doi-asserted-by":"publisher","DOI":"10.1038\/nature15535"},{"issue":"1","key":"2023061611200760000_2023.06.13.544841v1.4","first-page":"20","article-title":"Vector biology meets disease control: using basic research to fight vector-borne diseases","volume":"4","year":"2019","journal-title":"Nature Microbiology"},{"key":"2023061611200760000_2023.06.13.544841v1.5","doi-asserted-by":"publisher","DOI":"10.1016\/j.pt.2015.11.010"},{"issue":"1","key":"2023061611200760000_2023.06.13.544841v1.6","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1186\/1756-3305-7-76","article-title":"An online tool for mapping insecticide resistance in major Anopheles vectors of human malaria parasites and review of resistance status for the Afrotropical region","volume":"7","year":"2014","journal-title":"Parasites & Vectors"},{"key":"2023061611200760000_2023.06.13.544841v1.7","doi-asserted-by":"publisher","DOI":"10.3201\/eid1809.120262"},{"key":"2023061611200760000_2023.06.13.544841v1.8","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1186\/s12936-015-0847-4","article-title":"Characterizing the insecticide resistance of Anopheles gambiae in Mali","volume":"14","year":"2015","journal-title":"Malaria journal"},{"key":"2023061611200760000_2023.06.13.544841v1.9","doi-asserted-by":"publisher","DOI":"10.1016\/0169-4758(87)90196-7"},{"key":"2023061611200760000_2023.06.13.544841v1.10","doi-asserted-by":"crossref","unstructured":"Klassen W , Curtis CF . History of the Sterile Insect Technique. In: Dyck VA , Hendrichs J , Robinson AS , editors. Sterile Insect Technique: Principles and Practice in Area-Wide Integrated Pest Management. Dordrecht: Springer Netherlands; 2005. p. 3\u201336.","DOI":"10.1007\/1-4020-4051-2_1"},{"issue":"1","key":"2023061611200760000_2023.06.13.544841v1.11","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1111\/j.1570-7458.2012.01312.x","article-title":"Female mating failure and the failure of \u2018mating\u2019in sterile insect programs","volume":"146","year":"2013","journal-title":"Entomologia Experimentalis et Applicata"},{"key":"2023061611200760000_2023.06.13.544841v1.12","unstructured":"LaChance LE . The induction of dominant lethal mutations in insects by ionizing radiation and chemicals-as related to the sterile male technique of insect control. 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