Targeted Mutagenesis of Duplicated Genes in Soybean with Zinc-Finger Nucleases
We performed targeted mutagenesis of a transgene and nine endogenous soybean (Glycine max) genes using zinc-finger nucleases (ZFNs). A suite of ZFNs were engineered by the recently described context-dependent assembly platform—a rapid, open-source method for generating zinc-finger arrays. Specific Z...
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| Veröffentlicht in: | Plant physiology (Bethesda) 2011-06, Vol.156 (2), p.466-473 |
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| creator | Curtin, Shaun J. Zhang, Feng Sander, Jeffry D. Haun, William J. Starker, Colby Baltes, Nicholas J. Reyon, Deepak Dahlborg, Elizabeth J. Goodwin, Mathew J. Coffman, Andrew P. Dobbs, Drena Joung, J. Keith Voytas, Daniel F. Stupar, Robert M. |
| description | We performed targeted mutagenesis of a transgene and nine endogenous soybean (Glycine max) genes using zinc-finger nucleases (ZFNs). A suite of ZFNs were engineered by the recently described context-dependent assembly platform—a rapid, open-source method for generating zinc-finger arrays. Specific ZFNs targeting DICER-LIKE (DCL) genes and other genes involved in RNA silencing were cloned into a vector under an estrogen-inducible promoter. A hairy-root transformation system was employed to investigate the efficiency of ZFN mutagenesis at each target locus. Transgenic roots exhibited somatic mutations localized at the ZFN target sites for seven out of nine targeted genes. We next introduced a ZFN into soybean via whole-plant transformation and generated independent mutations in the paralogous genes DCL4a and DCL4b. The dcl4b mutation showed efficient heritable transmission of the ZFN-induced mutation in the subsequent generation. These findings indicate that ZFN-based mutagenesis provides an efficient method for making mutations in duplicate genes that are otherwise difficult to study due to redundancy. We also developed a publicly accessible Web-based tool to identify sites suitable for engineering context-dependent assembly ZFNs in the soybean genome. |
| doi_str_mv | 10.1104/pp.111.172981 |
| format | Article |
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Keith ; Voytas, Daniel F. ; Stupar, Robert M.</creator><creatorcontrib>Curtin, Shaun J. ; Zhang, Feng ; Sander, Jeffry D. ; Haun, William J. ; Starker, Colby ; Baltes, Nicholas J. ; Reyon, Deepak ; Dahlborg, Elizabeth J. ; Goodwin, Mathew J. ; Coffman, Andrew P. ; Dobbs, Drena ; Joung, J. Keith ; Voytas, Daniel F. ; Stupar, Robert M.</creatorcontrib><description>We performed targeted mutagenesis of a transgene and nine endogenous soybean (Glycine max) genes using zinc-finger nucleases (ZFNs). A suite of ZFNs were engineered by the recently described context-dependent assembly platform—a rapid, open-source method for generating zinc-finger arrays. Specific ZFNs targeting DICER-LIKE (DCL) genes and other genes involved in RNA silencing were cloned into a vector under an estrogen-inducible promoter. A hairy-root transformation system was employed to investigate the efficiency of ZFN mutagenesis at each target locus. Transgenic roots exhibited somatic mutations localized at the ZFN target sites for seven out of nine targeted genes. We next introduced a ZFN into soybean via whole-plant transformation and generated independent mutations in the paralogous genes DCL4a and DCL4b. The dcl4b mutation showed efficient heritable transmission of the ZFN-induced mutation in the subsequent generation. These findings indicate that ZFN-based mutagenesis provides an efficient method for making mutations in duplicate genes that are otherwise difficult to study due to redundancy. We also developed a publicly accessible Web-based tool to identify sites suitable for engineering context-dependent assembly ZFNs in the soybean genome.</description><identifier>ISSN: 0032-0889</identifier><identifier>ISSN: 1532-2548</identifier><identifier>EISSN: 1532-2548</identifier><identifier>DOI: 10.1104/pp.111.172981</identifier><identifier>PMID: 21464476</identifier><identifier>CODEN: PPHYA5</identifier><language>eng</language><publisher>Rockville, MD: American Society of Plant Biologists</publisher><subject>Base Sequence ; Biological and medical sciences ; BREAKTHROUGH TECHNOLOGIES ; Endonucleases - chemistry ; Endonucleases - metabolism ; Fundamental and applied biological sciences. Psychology ; Genes ; Genes, Duplicate - genetics ; Genes, Plant - genetics ; Genetic engineering ; Genetic mutation ; Genetic Techniques ; Genomes ; Glycine max - genetics ; Green Fluorescent Proteins - metabolism ; Inheritance Patterns - genetics ; Internet ; Molecular Sequence Data ; Mutagenesis ; Mutagenesis - genetics ; Mutation - genetics ; Plant physiology and development ; Plant Roots - genetics ; Plants ; Polymerase Chain Reaction ; Soybeans ; Transgenes ; Transgenes - genetics ; Zinc ; Zinc Fingers - genetics</subject><ispartof>Plant physiology (Bethesda), 2011-06, Vol.156 (2), p.466-473</ispartof><rights>2011 American Society of Plant Biologists</rights><rights>2015 INIST-CNRS</rights><rights>2011 American Society of Plant Biologists 2011</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c438t-7c36c2cd9f6b99b3ce02150ec056e77deba89739e0df05968615589dd12d2da83</citedby><cites>FETCH-LOGICAL-c438t-7c36c2cd9f6b99b3ce02150ec056e77deba89739e0df05968615589dd12d2da83</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/41434317$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/41434317$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,776,780,799,881,27901,27902,57992,58225</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=24257592$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21464476$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Curtin, Shaun J.</creatorcontrib><creatorcontrib>Zhang, Feng</creatorcontrib><creatorcontrib>Sander, Jeffry D.</creatorcontrib><creatorcontrib>Haun, William J.</creatorcontrib><creatorcontrib>Starker, Colby</creatorcontrib><creatorcontrib>Baltes, Nicholas J.</creatorcontrib><creatorcontrib>Reyon, Deepak</creatorcontrib><creatorcontrib>Dahlborg, Elizabeth J.</creatorcontrib><creatorcontrib>Goodwin, Mathew J.</creatorcontrib><creatorcontrib>Coffman, Andrew P.</creatorcontrib><creatorcontrib>Dobbs, Drena</creatorcontrib><creatorcontrib>Joung, J. Keith</creatorcontrib><creatorcontrib>Voytas, Daniel F.</creatorcontrib><creatorcontrib>Stupar, Robert M.</creatorcontrib><title>Targeted Mutagenesis of Duplicated Genes in Soybean with Zinc-Finger Nucleases</title><title>Plant physiology (Bethesda)</title><addtitle>Plant Physiol</addtitle><description>We performed targeted mutagenesis of a transgene and nine endogenous soybean (Glycine max) genes using zinc-finger nucleases (ZFNs). A suite of ZFNs were engineered by the recently described context-dependent assembly platform—a rapid, open-source method for generating zinc-finger arrays. Specific ZFNs targeting DICER-LIKE (DCL) genes and other genes involved in RNA silencing were cloned into a vector under an estrogen-inducible promoter. A hairy-root transformation system was employed to investigate the efficiency of ZFN mutagenesis at each target locus. Transgenic roots exhibited somatic mutations localized at the ZFN target sites for seven out of nine targeted genes. We next introduced a ZFN into soybean via whole-plant transformation and generated independent mutations in the paralogous genes DCL4a and DCL4b. The dcl4b mutation showed efficient heritable transmission of the ZFN-induced mutation in the subsequent generation. These findings indicate that ZFN-based mutagenesis provides an efficient method for making mutations in duplicate genes that are otherwise difficult to study due to redundancy. We also developed a publicly accessible Web-based tool to identify sites suitable for engineering context-dependent assembly ZFNs in the soybean genome.</description><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>BREAKTHROUGH TECHNOLOGIES</subject><subject>Endonucleases - chemistry</subject><subject>Endonucleases - metabolism</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genes</subject><subject>Genes, Duplicate - genetics</subject><subject>Genes, Plant - genetics</subject><subject>Genetic engineering</subject><subject>Genetic mutation</subject><subject>Genetic Techniques</subject><subject>Genomes</subject><subject>Glycine max - genetics</subject><subject>Green Fluorescent Proteins - metabolism</subject><subject>Inheritance Patterns - genetics</subject><subject>Internet</subject><subject>Molecular Sequence Data</subject><subject>Mutagenesis</subject><subject>Mutagenesis - genetics</subject><subject>Mutation - genetics</subject><subject>Plant physiology and development</subject><subject>Plant Roots - genetics</subject><subject>Plants</subject><subject>Polymerase Chain Reaction</subject><subject>Soybeans</subject><subject>Transgenes</subject><subject>Transgenes - genetics</subject><subject>Zinc</subject><subject>Zinc Fingers - genetics</subject><issn>0032-0889</issn><issn>1532-2548</issn><issn>1532-2548</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkDlPxDAQhS0EguUoKUFpKAMeH7HdICFuiaMAGprIsSeLUUgiOwvi35PVwgLVs-Z988Z6hOwCPQSg4qjvR4VDUMxoWCETkJzlTAq9SiaUjm-qtdkgmym9UkqBg1gnGwxEIYQqJuTu0cYpDuiz29lgp9hiCinr6uxs1jfB2blzOZ9moc0eus8KbZt9hOElew6tyy9CO8WY3c1cgzZh2iZrtW0S7nzrFnm6OH88vcpv7i-vT09ucie4HnLleOGY86YuKmMq7pAykBQdlQUq5bGy2ihukPqaSlPoAqTUxntgnnmr-RY5XuT2s-oNvcN2iLYp-xjebPwsOxvK_04bXspp915yUIpJOgbkiwAXu5Qi1stdoOW82LLvR4VyUezI7_89uKR_mhyBg2_AJmebOtrWhfTLCSaVNGzk9hbcaxq6uPQFCC7Gz_EvM6OLdg</recordid><startdate>20110601</startdate><enddate>20110601</enddate><creator>Curtin, Shaun J.</creator><creator>Zhang, Feng</creator><creator>Sander, Jeffry D.</creator><creator>Haun, William J.</creator><creator>Starker, Colby</creator><creator>Baltes, Nicholas J.</creator><creator>Reyon, Deepak</creator><creator>Dahlborg, Elizabeth J.</creator><creator>Goodwin, Mathew J.</creator><creator>Coffman, Andrew P.</creator><creator>Dobbs, Drena</creator><creator>Joung, J. 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Psychology</topic><topic>Genes</topic><topic>Genes, Duplicate - genetics</topic><topic>Genes, Plant - genetics</topic><topic>Genetic engineering</topic><topic>Genetic mutation</topic><topic>Genetic Techniques</topic><topic>Genomes</topic><topic>Glycine max - genetics</topic><topic>Green Fluorescent Proteins - metabolism</topic><topic>Inheritance Patterns - genetics</topic><topic>Internet</topic><topic>Molecular Sequence Data</topic><topic>Mutagenesis</topic><topic>Mutagenesis - genetics</topic><topic>Mutation - genetics</topic><topic>Plant physiology and development</topic><topic>Plant Roots - genetics</topic><topic>Plants</topic><topic>Polymerase Chain Reaction</topic><topic>Soybeans</topic><topic>Transgenes</topic><topic>Transgenes - genetics</topic><topic>Zinc</topic><topic>Zinc Fingers - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Curtin, Shaun J.</creatorcontrib><creatorcontrib>Zhang, Feng</creatorcontrib><creatorcontrib>Sander, Jeffry D.</creatorcontrib><creatorcontrib>Haun, William J.</creatorcontrib><creatorcontrib>Starker, Colby</creatorcontrib><creatorcontrib>Baltes, Nicholas J.</creatorcontrib><creatorcontrib>Reyon, Deepak</creatorcontrib><creatorcontrib>Dahlborg, Elizabeth J.</creatorcontrib><creatorcontrib>Goodwin, Mathew J.</creatorcontrib><creatorcontrib>Coffman, Andrew P.</creatorcontrib><creatorcontrib>Dobbs, Drena</creatorcontrib><creatorcontrib>Joung, J. 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Keith</au><au>Voytas, Daniel F.</au><au>Stupar, Robert M.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Targeted Mutagenesis of Duplicated Genes in Soybean with Zinc-Finger Nucleases</atitle><jtitle>Plant physiology (Bethesda)</jtitle><addtitle>Plant Physiol</addtitle><date>2011-06-01</date><risdate>2011</risdate><volume>156</volume><issue>2</issue><spage>466</spage><epage>473</epage><pages>466-473</pages><issn>0032-0889</issn><issn>1532-2548</issn><eissn>1532-2548</eissn><coden>PPHYA5</coden><abstract>We performed targeted mutagenesis of a transgene and nine endogenous soybean (Glycine max) genes using zinc-finger nucleases (ZFNs). A suite of ZFNs were engineered by the recently described context-dependent assembly platform—a rapid, open-source method for generating zinc-finger arrays. Specific ZFNs targeting DICER-LIKE (DCL) genes and other genes involved in RNA silencing were cloned into a vector under an estrogen-inducible promoter. A hairy-root transformation system was employed to investigate the efficiency of ZFN mutagenesis at each target locus. Transgenic roots exhibited somatic mutations localized at the ZFN target sites for seven out of nine targeted genes. We next introduced a ZFN into soybean via whole-plant transformation and generated independent mutations in the paralogous genes DCL4a and DCL4b. The dcl4b mutation showed efficient heritable transmission of the ZFN-induced mutation in the subsequent generation. These findings indicate that ZFN-based mutagenesis provides an efficient method for making mutations in duplicate genes that are otherwise difficult to study due to redundancy. We also developed a publicly accessible Web-based tool to identify sites suitable for engineering context-dependent assembly ZFNs in the soybean genome.</abstract><cop>Rockville, MD</cop><pub>American Society of Plant Biologists</pub><pmid>21464476</pmid><doi>10.1104/pp.111.172981</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| source | Jstor Complete Legacy; Oxford University Press Journals All Titles (1996-Current); MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals |
| subjects | Base Sequence Biological and medical sciences BREAKTHROUGH TECHNOLOGIES Endonucleases - chemistry Endonucleases - metabolism Fundamental and applied biological sciences. Psychology Genes Genes, Duplicate - genetics Genes, Plant - genetics Genetic engineering Genetic mutation Genetic Techniques Genomes Glycine max - genetics Green Fluorescent Proteins - metabolism Inheritance Patterns - genetics Internet Molecular Sequence Data Mutagenesis Mutagenesis - genetics Mutation - genetics Plant physiology and development Plant Roots - genetics Plants Polymerase Chain Reaction Soybeans Transgenes Transgenes - genetics Zinc Zinc Fingers - genetics |
| title | Targeted Mutagenesis of Duplicated Genes in Soybean with Zinc-Finger Nucleases |
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