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In this study, we aimed to develop high\u2010oleic soybean (\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Glycine max<\/jats:italic>\n                    <\/jats:styled-content>\n                    ) by simultaneously targeting fatty acid desaturase 2 (\n                    <jats:italic>FAD2<\/jats:italic>\n                    ) and fatty acyl\u2010ACP thioesterase B (\n                    <jats:italic>FATB<\/jats:italic>\n                    ) gene families using CRISPR\/Cas9. Considering the paleopolyploid genome of soybean, multiple sgRNAs were designed to target\n                    <jats:italic>GmFAD2\u20101<\/jats:italic>\n                    ,\n                    <jats:italic>GmFAD2\u20102<\/jats:italic>\n                    , and\n                    <jats:italic>GmFATB<\/jats:italic>\n                    genes expressed during seed development. Compared with targeting\n                    <jats:italic>GmFAD2\u20101<\/jats:italic>\n                    alone (~83%), additional editing of\n                    <jats:italic>GmFAD2\u20102B, GmFAD2\u20102C,<\/jats:italic>\n                    and\n                    <jats:italic>GmFATB1a<\/jats:italic>\n                    increased the 18:1 fatty acid content to over 90%. Editing of\n                    <jats:italic>GmFATB<\/jats:italic>\n                    significantly reduced the SFA content by more than 40% compared with wild\u2010type (WT). Reduced absolute SFA content was also observed in GmFAD2\u2010only edited lines with decreased\n                    <jats:italic>GmFATB<\/jats:italic>\n                    expression. Notably, all evaluated high\u2010oleic genotypes in the Williams82\u2010background showed higher mean total fatty acid (TFA) content than the WT, both per unit seed mass and per seed. Correlation analysis of 805 individual seed profiles from Williams82\u2010background lines further characterized the relationship between 18:1 fatty acid composition and TFA content per unit seed mass. Gene expression analysis revealed no consistent increase in triacylglycerol (TAG) synthesis genes, whereas the expression of\n                    <jats:italic>sugar\u2010dependent 1<\/jats:italic>\n                    (\n                    <jats:italic>SDP1<\/jats:italic>\n                    ) was reduced. In contrast, genes involved in phosphatidic acid (PA) metabolism, including\n                    <jats:italic>diacylglycerol kinase<\/jats:italic>\n                    (\n                    <jats:italic>DGK<\/jats:italic>\n                    ) and\n                    <jats:italic>phospholipase D<\/jats:italic>\n                    (\n                    <jats:italic>PLD<\/jats:italic>\n                    ), were partially upregulated. These transcriptional changes support a hypothesis that altered lipid turnover may contribute to the higher TFA phenotype. Collectively, this study defines effective multiplex target combinations for ultra\u2010high oleic, low\u2010saturated soybean.\n                  <\/jats:p>","DOI":"10.1111\/pbi.70758","type":"journal-article","created":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T06:08:01Z","timestamp":1789452481000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Multiplex\n                    <i>FAD2\/FATB<\/i>\n                    Editing Generates Ultra\u2010High\u2010Oleic, Low\u2010Saturate Soybean With Increased Seed Fatty Acid Content"],"prefix":"10.1111","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8685-5223","authenticated-orcid":false,"given":"Won Nyeong","family":"Kim","sequence":"first","affiliation":[{"name":"Department of Bioindustry and Bioresource Engineering Sejong University  Seoul Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1671-1881","authenticated-orcid":false,"given":"Yu\u2010Ri","family":"Choi","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology Sejong University  Seoul South Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hye Jeong","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Biomaterials Engineering, College of Natural Resources and Life Sciences Dong\u2010A University  Busan Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Young\u2010Soo","family":"Chung","sequence":"additional","affiliation":[{"name":"Department of Biomaterials Engineering, College of Natural Resources and Life Sciences Dong\u2010A University  Busan Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4566-3057","authenticated-orcid":false,"given":"Hyun Uk","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Bioindustry and Bioresource Engineering Sejong University  Seoul Republic of Korea"},{"name":"Department of Molecular Biology Sejong University  Seoul South Korea"},{"name":"Institute for Advanced Plant Breeding and Phytochemicals (IAPBP) Sejong University  Seoul Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,9,14]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1270\/jsbbs.58.447"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.3389\/fpls.2013.00001"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.2174\/1875036201307010001"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1111\/tpj.14195"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.heliyon.2024.e34669"},{"key":"e_1_2_10_7_1","doi-asserted-by":"crossref","DOI":"10.1186\/s12896-024-00936-4","article-title":"Genome\u2010Wide Identification and Diversity of FAD2, FAD3 and FAE1 Genes in Terms of Biotechnological Importance in Camelina Species","volume":"24","author":"Blume R. 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