Site‐Specific Gene Targeting Using Transcription Activator‐Like Effector (TALE)‐Based Nuclease in Brassica oleracea
Identifieur interne : 002106 ( Main/Exploration ); précédent : 002105; suivant : 002107Site‐Specific Gene Targeting Using Transcription Activator‐Like Effector (TALE)‐Based Nuclease in Brassica oleracea
Auteurs : Zijian Sun [République populaire de Chine] ; Nianzu Li [République populaire de Chine] ; Guodong Huang [République populaire de Chine] ; Junqiang Xu [République populaire de Chine] ; Yu Pan [République populaire de Chine] ; Zhimin Wang [République populaire de Chine] ; Qinglin Tang [République populaire de Chine] ; Ming Song [République populaire de Chine] ; Xiaojia Wang [République populaire de Chine]Source :
- Journal of Integrative Plant Biology [ 1672-9072 ] ; 2013-11.
English descriptors
- Teeft :
- Amino, Amino acid, Amino acid residues, Assay, Backbone vector, Bglii linearized plasmid, Binary vector, Biol, Biotechnol, Boch, Brassica, Brassica oleracea, Cleavage, Cleavage activity, Clone, Deletion, Digestion, Dsbs, Ebes, Ecori site, Effector, Endonuclease, Expression cassette, Foki, Foki cleavage domain, Foki domain, Frigida, Gene, Genome, Genome editing, Golden gate, Hybrid, Hybrid nuclease, Infection medium, Insertion, Integrative plant biology, Joung, Mahfouz, Module, Morbitzer, Mutant, Mutation, Nger, Nger nucleases, Nuclease, Nucleic acids, Nucleotide, Oleracea, Plant biol, Plant biotechnol, Plant cells, Plant physiol, Plasmid, Previous studies, Primer, Restriction enzymes, Sequence analysis, Sequencing, Spacer, Talen, Talen pair, Talen pairs, Talens, Target sequence, Target site, Transcription, Transcription effector, Transcription effector nuclease, Transient assay, Uida, Uida activity, Unit assembly, Voytas, Wang, Wild type, Zhang.
Abstract
Site‐specific recognition modules with DNA nuclease have tremendous potential as molecular tools for genome targeting. The type III transcription activator‐like effectors (TALEs) contain a DNA binding domain consisting of tandem repeats that can be engineered to bind user‐defined specific DNA sequences. We demonstrated that customized TALE‐based nucleases (TALENs), constructed using a method called “unit assembly”, specifically target the endogenous FRIGIDA gene in Brassica oleracea L. var. capitata L. The results indicate that the TALENs bound to the target site and cleaved double‐strand DNA in vitro and in vivo, whereas the effector binding elements have a 23 bp spacer. The T7 endonuclease I assay and sequencing data show that TALENs made double‐strand breaks, which were repaired by a non‐homologous end‐joining pathway within the target sequence. These data show the feasibility of applying customized TALENs to target and modify the genome with deletions in those organisms that are still in lacking gene target methods to provide germplasms in breeding improvement.
Url:
DOI: 10.1111/jipb.12091
Affiliations:
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Horticulture Science for Southern Mountainous Regions, Southwest University, 400715, Chongqing</wicri:regionArea><wicri:noRegion>Chongqing</wicri:noRegion></affiliation></author><author><name sortKey="Pan, Yu" sort="Pan, Yu" uniqKey="Pan Y" first="Yu" last="Pan">Yu Pan</name><affiliation wicri:level="1"><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Horticulture Science for Southern Mountainous Regions, Southwest University, 400715, Chongqing</wicri:regionArea><wicri:noRegion>Chongqing</wicri:noRegion></affiliation></author><author><name sortKey="Wang, Zhimin" sort="Wang, Zhimin" uniqKey="Wang Z" first="Zhimin" last="Wang">Zhimin Wang</name><affiliation wicri:level="1"><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Key Laboratory of Horticulture Science for Southern Mountainous Regions, Southwest University, 400715, 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from="1092">1092</biblScope><biblScope unit="page" to="1103">1103</biblScope><biblScope unit="page-count">12</biblScope><date type="published" when="2013-11">2013-11</date></imprint><idno type="ISSN">1672-9072</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">1672-9072</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Amino</term><term>Amino acid</term><term>Amino acid residues</term><term>Assay</term><term>Backbone vector</term><term>Bglii linearized plasmid</term><term>Binary vector</term><term>Biol</term><term>Biotechnol</term><term>Boch</term><term>Brassica</term><term>Brassica oleracea</term><term>Cleavage</term><term>Cleavage activity</term><term>Clone</term><term>Deletion</term><term>Digestion</term><term>Dsbs</term><term>Ebes</term><term>Ecori site</term><term>Effector</term><term>Endonuclease</term><term>Expression cassette</term><term>Foki</term><term>Foki cleavage domain</term><term>Foki domain</term><term>Frigida</term><term>Gene</term><term>Genome</term><term>Genome editing</term><term>Golden gate</term><term>Hybrid</term><term>Hybrid nuclease</term><term>Infection medium</term><term>Insertion</term><term>Integrative plant biology</term><term>Joung</term><term>Mahfouz</term><term>Module</term><term>Morbitzer</term><term>Mutant</term><term>Mutation</term><term>Nger</term><term>Nger nucleases</term><term>Nuclease</term><term>Nucleic acids</term><term>Nucleotide</term><term>Oleracea</term><term>Plant biol</term><term>Plant biotechnol</term><term>Plant cells</term><term>Plant physiol</term><term>Plasmid</term><term>Previous studies</term><term>Primer</term><term>Restriction enzymes</term><term>Sequence analysis</term><term>Sequencing</term><term>Spacer</term><term>Talen</term><term>Talen pair</term><term>Talen pairs</term><term>Talens</term><term>Target sequence</term><term>Target site</term><term>Transcription</term><term>Transcription effector</term><term>Transcription effector nuclease</term><term>Transient assay</term><term>Uida</term><term>Uida activity</term><term>Unit assembly</term><term>Voytas</term><term>Wang</term><term>Wild type</term><term>Zhang</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Site‐specific recognition modules with DNA nuclease have tremendous potential as molecular tools for genome targeting. The type III transcription activator‐like effectors (TALEs) contain a DNA binding domain consisting of tandem repeats that can be engineered to bind user‐defined specific DNA sequences. We demonstrated that customized TALE‐based nucleases (TALENs), constructed using a method called “unit assembly”, specifically target the endogenous FRIGIDA gene in Brassica oleracea L. var. capitata L. The results indicate that the TALENs bound to the target site and cleaved double‐strand DNA in vitro and in vivo, whereas the effector binding elements have a 23 bp spacer. The T7 endonuclease I assay and sequencing data show that TALENs made double‐strand breaks, which were repaired by a non‐homologous end‐joining pathway within the target sequence. These data show the feasibility of applying customized TALENs to target and modify the genome with deletions in those organisms that are still in lacking gene target methods to provide germplasms in breeding improvement.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country></list><tree><country name="République populaire de Chine"><noRegion><name sortKey="Sun, Zijian" sort="Sun, Zijian" uniqKey="Sun Z" first="Zijian" last="Sun">Zijian Sun</name></noRegion><name sortKey="Huang, Guodong" sort="Huang, Guodong" uniqKey="Huang G" first="Guodong" last="Huang">Guodong Huang</name><name sortKey="Li, Nianzu" sort="Li, Nianzu" uniqKey="Li N" first="Nianzu" last="Li">Nianzu Li</name><name sortKey="Pan, Yu" sort="Pan, Yu" uniqKey="Pan Y" first="Yu" last="Pan">Yu Pan</name><name sortKey="Song, Ming" sort="Song, Ming" uniqKey="Song M" first="Ming" last="Song">Ming Song</name><name sortKey="Song, Ming" sort="Song, Ming" uniqKey="Song M" first="Ming" last="Song">Ming Song</name><name sortKey="Tang, Qinglin" sort="Tang, Qinglin" uniqKey="Tang Q" first="Qinglin" last="Tang">Qinglin Tang</name><name sortKey="Wang, Xiaojia" sort="Wang, Xiaojia" uniqKey="Wang X" first="Xiaojia" last="Wang">Xiaojia Wang</name><name sortKey="Wang, Xiaojia" sort="Wang, Xiaojia" uniqKey="Wang X" first="Xiaojia" last="Wang">Xiaojia Wang</name><name sortKey="Wang, Zhimin" sort="Wang, Zhimin" uniqKey="Wang Z" first="Zhimin" last="Wang">Zhimin Wang</name><name sortKey="Xu, Junqiang" sort="Xu, Junqiang" uniqKey="Xu J" first="Junqiang" last="Xu">Junqiang Xu</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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