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Precise gene editing techniques were used to generate Sal1 mutants in hexaploid bread wheat. The CRISPR (Clustered Regulatory Interspaced Short Palindromic Repeats) Cas9 system with three guide RNAs (gRNAs) was used to inactivate six Sal1 homologous genes within the Bobwhite wheat genome. The resulting mutant wheat plants with all their Sal1 genes disabled had slimmer stems, had a modest reduction in biomass and senesced more slowly in water limiting conditions, but did not exhibit improved yield under drought conditions. Our results show that multiplexed gRNAs enabled effective targeted gene editing of the Sal1 gene family in hexaploid wheat. These Sal1 mutant wheat plants will be a resource for further research studying the function of this gene family in wheat.<\/jats:p>","DOI":"10.3390\/plants11172259","type":"journal-article","created":{"date-parts":[[2022,8,31]],"date-time":"2022-08-31T00:13:56Z","timestamp":1661904836000},"page":"2259","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":43,"title":["CRISPR-Cas9 Gene Editing of the Sal1 Gene Family in Wheat"],"prefix":"10.3390","volume":"11","author":[{"given":"Toni","family":"Mohr","sequence":"first","affiliation":[{"name":"USDA-ARS, Crop Improvement and Genetics Unit, Albany, CA 94710, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James","family":"Horstman","sequence":"additional","affiliation":[{"name":"USDA-ARS, Crop Improvement and Genetics Unit, Albany, CA 94710, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yong Q.","family":"Gu","sequence":"additional","affiliation":[{"name":"USDA-ARS, Crop Improvement and Genetics Unit, Albany, CA 94710, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1113-3581","authenticated-orcid":false,"given":"Nagwa I.","family":"Elarabi","sequence":"additional","affiliation":[{"name":"Department of Genetics, Faculty of Agriculture, Cairo University, Giza 12613, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9290-5944","authenticated-orcid":false,"given":"Naglaa A.","family":"Abdallah","sequence":"additional","affiliation":[{"name":"Department of Genetics, Faculty of Agriculture, Cairo University, Giza 12613, Egypt"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6591-6699","authenticated-orcid":false,"given":"Roger","family":"Thilmony","sequence":"additional","affiliation":[{"name":"USDA-ARS, Crop Improvement and Genetics Unit, Albany, CA 94710, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,8,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1537","DOI":"10.1093\/jxb\/erp058","article-title":"Wheat","volume":"60","author":"Shewry","year":"2009","journal-title":"J. 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