{"title":"拟南芥质体基因组中的靶向C-to-T碱基编辑","authors":"Issei Nakazato, Shin-ichi Arimura","doi":"10.1002/cpz1.70075","DOIUrl":null,"url":null,"abstract":"<p><i>Arabidopsis thaliana</i>, particularly the ecotype Columbia-0 (Col-0), has been extensively employed in the study of genetics of the nuclear genome. However, the difficulty of modifying the plastid genome of Col-0, the most widely used ecotype, has hindered investigation of the functional interactions between nuclear-encoded and plastid-encoded genes in this ecotype. Recently, we achieved targeted base editing, substituting a specific C:G pair with a T:A pair in the plastid genome of Col-0 through the application of genome-editing technology. This article introduces the method employed to accomplish this targeted base editing. The process involves four steps: (i) designing and constructing a binary vector encoding the genome-editing enzyme, (ii) introducing the binary vector into the nuclear genome of Col-0 via floral dipping, (iii) identifying base-edited plants, and (iv) verifying inheritance of the edited base(s) by the next generation. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Design and construction of a binary vector encoding ptpTALECD or ptpTALECD_v2</p><p><b>Basic Protocol 2</b>: <i>Agrobacterium</i>-mediated introduction of a binary vector into the <i>Arabidopsis</i> nuclear genome</p><p><b>Basic Protocol 3</b>: Selection of plants harboring T-DNA in the nucleus and detection of base editing in the plastid genome</p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":"5 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2025-01-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cpz1.70075","citationCount":"0","resultStr":"{\"title\":\"Targeted C-to-T Base Editing in the Arabidopsis Plastid Genome\",\"authors\":\"Issei Nakazato, Shin-ichi Arimura\",\"doi\":\"10.1002/cpz1.70075\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><i>Arabidopsis thaliana</i>, particularly the ecotype Columbia-0 (Col-0), has been extensively employed in the study of genetics of the nuclear genome. However, the difficulty of modifying the plastid genome of Col-0, the most widely used ecotype, has hindered investigation of the functional interactions between nuclear-encoded and plastid-encoded genes in this ecotype. Recently, we achieved targeted base editing, substituting a specific C:G pair with a T:A pair in the plastid genome of Col-0 through the application of genome-editing technology. This article introduces the method employed to accomplish this targeted base editing. The process involves four steps: (i) designing and constructing a binary vector encoding the genome-editing enzyme, (ii) introducing the binary vector into the nuclear genome of Col-0 via floral dipping, (iii) identifying base-edited plants, and (iv) verifying inheritance of the edited base(s) by the next generation. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Design and construction of a binary vector encoding ptpTALECD or ptpTALECD_v2</p><p><b>Basic Protocol 2</b>: <i>Agrobacterium</i>-mediated introduction of a binary vector into the <i>Arabidopsis</i> nuclear genome</p><p><b>Basic Protocol 3</b>: Selection of plants harboring T-DNA in the nucleus and detection of base editing in the plastid genome</p>\",\"PeriodicalId\":93970,\"journal\":{\"name\":\"Current protocols\",\"volume\":\"5 1\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2025-01-06\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cpz1.70075\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Current protocols\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/cpz1.70075\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current protocols","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpz1.70075","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0