\n
\"An improved recombineering toolset for plants\"<\/strong><\/p>\nAuthors:<\/em> Javier Brumos, Chengsong Zhao, Yan Gong, David Soriano, Arjun P. Patel, Miguel A Perez-Amador, Anna N Stepanova and Jose M Alonso, North Carolina State University<\/p>\nPublished:<\/em> Oct. 30, The Plant Cell<\/em><\/p>\nDOI:<\/em> 10.1105\/tpc.19.00431<\/p>\nAbstract:<\/strong> Gene functional studies often rely on the expression of a gene of interest as transcriptional and translational fusions with specialized tags. Ideally, this is done in the native chromosomal contexts to avoid potential misexpression artifacts. Although recent improvements in genome editing make it possible to directly modify the target genes in their native chromosomal location, classical transgenesis is still the preferred experimental approach chosen in most gene tagging studies because of its time efficiency and accessibility. We have developed a recombineering-based tagging system that brings together the convenience of the classical transgenic approaches and the high degree of confidence in the obtained results provided by the direct chromosomal tagging achievable by genome editing strategies. These simple, scalable and customizable recombineering toolsets and protocols allow for the generation of a variety of genetic modifications. In addition, a highly efficient recombinase-mediated cassette exchange system has been developed to facilitate the transfer of the desired sequences from a BAC clone to a transformation-compatible binary vector, expanding the use of the recombineering approaches beyond Arabidopsis. The utility of this system is demonstrated by the generation of over 250 whole-gene translational fusions and 123 Arabidopsis transgenic lines corresponding to 62 auxin-related genes, and the characterization of the translational reporter expression patterns for 14 auxin biosynthesis genes.<\/p>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<\/div>"},"excerpt":{"rendered":"NC State researchers share a new set of genetic engineering tools that makes modifying large plant genes faster and easier. This should improve the precision and reliability of the foundation for innovations in pest resistance, yield increases and growing season extensions. <\/p>\n","protected":false},"author":2360,"featured_media":173567,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"source":"","ncst_custom_author":"","ncst_show_custom_author":false,"ncst_dynamicHeaderBlockName":"ncst\/default-post-header","ncst_dynamicHeaderData":"{\"showAuthor\":true,\"showDate\":true,\"showFeaturedVideo\":false,\"caption\":\"Anna Stepanova and Jose Alonso shared a new genetic engineering toolset that makes working with large plant genes easier and faster with the plant science community.\",\"displayCategoryID\":1171}","ncst_content_audit_freq":"","ncst_content_audit_date":"","footnotes":"","_links_to":"","_links_to_target":""},"categories":[1171,1633,1181,1163],"tags":[1863,338],"_ncst_magazine_issue":[],"coauthors":[1677],"displayCategory":{"term_id":1171,"name":"Faculty and Staff","slug":"faculty-and-staff","term_group":0,"term_taxonomy_id":1171,"taxonomy":"category","description":"","parent":0,"count":1094,"filter":"raw"},"acf":[],"_links":{"self":[{"href":"https:\/\/cals.ncsu.edu\/wp-json\/wp\/v2\/posts\/173566"}],"collection":[{"href":"https:\/\/cals.ncsu.edu\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/cals.ncsu.edu\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/cals.ncsu.edu\/wp-json\/wp\/v2\/users\/2360"}],"replies":[{"embeddable":true,"href":"https:\/\/cals.ncsu.edu\/wp-json\/wp\/v2\/comments?post=173566"}],"version-history":[{"count":5,"href":"https:\/\/cals.ncsu.edu\/wp-json\/wp\/v2\/posts\/173566\/revisions"}],"predecessor-version":[{"id":193671,"href":"https:\/\/cals.ncsu.edu\/wp-json\/wp\/v2\/posts\/173566\/revisions\/193671"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/cals.ncsu.edu\/wp-json\/wp\/v2\/media\/173567"}],"wp:attachment":[{"href":"https:\/\/cals.ncsu.edu\/wp-json\/wp\/v2\/media?parent=173566"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/cals.ncsu.edu\/wp-json\/wp\/v2\/categories?post=173566"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/cals.ncsu.edu\/wp-json\/wp\/v2\/tags?post=173566"},{"taxonomy":"_ncst_magazine_issue","embeddable":true,"href":"https:\/\/cals.ncsu.edu\/wp-json\/wp\/v2\/_ncst_magazine_issue?post=173566"},{"taxonomy":"author","embeddable":true,"href":"https:\/\/cals.ncsu.edu\/wp-json\/wp\/v2\/coauthors?post=173566"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}