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Most previous research has focused on the mechanistic understanding of biological processes in <jats:italic>An. sinensis<\/jats:italic> and novel ways of interrupting malaria transmission. However, the development of functional genomics and genetics\u2010based vector control strategies against <jats:italic>An. sinensis<\/jats:italic> remain limited because of insufficient site\u2010specific genome editing tools.<\/jats:p><\/jats:sec><jats:sec><jats:title>RESULTS<\/jats:title><jats:p>We report the first successful application of the CRISPR\/Cas9 mediated knock\u2010in for highly efficient, site\u2010specific mutagenesis in <jats:italic>An. sinensis<\/jats:italic>. The EGFP marker gene driven by the 3\u2009\u00d7\u2009P3 promoter was precisely integrated into the odorant receptor co\u2010receptor (<jats:italic>Orco<\/jats:italic>) by direct injections of Cas9 protein, double\u2010stranded DNA donor, and <jats:italic>Orco<\/jats:italic>\u2010gRNA. We achieved a mutation rate of 3.77%, similar to rates in other mosquito species. Precise knock\u2010in at the intended locus was confirmed by polymerase chain reaction (PCR) amplification and sequencing. The Orco mutation severely impaired mosquito sensitivity to some odors and their ability to locate and discriminate a human host.<\/jats:p><\/jats:sec><jats:sec><jats:title>CONCLUSION<\/jats:title><jats:p><jats:italic>Orco<\/jats:italic> was confirmed as a key mediator of multiple olfactory\u2010driven behaviors in the <jats:italic>An<\/jats:italic>. <jats:italic>sinensis<\/jats:italic> life cycle, highlighting the importance of <jats:italic>Orco<\/jats:italic> as a key molecular target for malaria control. The results also demonstrated that CRISPR\/Cas9 was a simple and highly efficient genome editing technique for <jats:italic>An. sinensis<\/jats:italic> and could be used to develop genetic control tools for this vector. \u00a9 2022 Society of Chemical Industry.<\/jats:p><\/jats:sec>","DOI":"10.1002\/ps.6954","type":"journal-article","created":{"date-parts":[[2022,4,29]],"date-time":"2022-04-29T01:44:33Z","timestamp":1651196673000},"page":"3294-3304","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["<scp>CRISPR\/Cas9<\/scp> mediates efficient site\u2010specific mutagenesis of the odorant receptor co\u2010receptor (<i>Orco<\/i>) in the malaria vector <i>Anopheles sinensis<\/i>"],"prefix":"10.1002","volume":"78","author":[{"given":"You","family":"Wang","sequence":"first","affiliation":[{"name":"Chongqing Key Laboratory of Vector Insects; Institute of Entomology and Molecular Biology Chongqing Normal University  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