ULtiMATE system for rapid assembly of customized TAL effectors.
Engineered TAL-effector nucleases (TALENs) and TALE-based constructs have become powerful tools for eukaryotic genome editing. Although many methods have been reported, it remains a challenge for the assembly of designer-based TALE repeats in a fast, precise and cost-effective manner. We present an...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2013-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3815405?pdf=render |
| _version_ | 1818701116823568384 |
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| author | Junjiao Yang Pengfei Yuan Dingqiao Wen Ying Sheng Shiyou Zhu Yuezhou Yu Xiang Gao Wensheng Wei |
| author_facet | Junjiao Yang Pengfei Yuan Dingqiao Wen Ying Sheng Shiyou Zhu Yuezhou Yu Xiang Gao Wensheng Wei |
| author_sort | Junjiao Yang |
| collection | DOAJ |
| description | Engineered TAL-effector nucleases (TALENs) and TALE-based constructs have become powerful tools for eukaryotic genome editing. Although many methods have been reported, it remains a challenge for the assembly of designer-based TALE repeats in a fast, precise and cost-effective manner. We present an ULtiMATE (USER-based Ligation Mediated Assembly of TAL Effector) system for speedy and accurate assembly of customized TALE constructs. This method takes advantage of uracil-specific excision reagent (USER) to create multiple distinct sticky ends between any neighboring DNA fragments for specific ligation. With pre-assembled templates, multiple TALE DNA-binding domains could be efficiently assembled in order within hours with minimal manual operation. This system has been demonstrated to produce both functional TALENs for effective gene knockout and TALE-mediated gene-specific transcription activation (TALE-TA). The feature of both ease-of-operation and high efficiency of ULtiMATE system makes it not only an ideal method for biologic labs, but also an approach well suited for large-scale assembly of TALENs and any other TALE-based constructions. |
| first_indexed | 2024-12-17T15:15:44Z |
| format | Article |
| id | doaj.art-1fe00fc8dc924c8ba1589ac19d8bc174 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-17T15:15:44Z |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-1fe00fc8dc924c8ba1589ac19d8bc1742022-12-21T21:43:32ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0189e7564910.1371/journal.pone.0075649ULtiMATE system for rapid assembly of customized TAL effectors.Junjiao YangPengfei YuanDingqiao WenYing ShengShiyou ZhuYuezhou YuXiang GaoWensheng WeiEngineered TAL-effector nucleases (TALENs) and TALE-based constructs have become powerful tools for eukaryotic genome editing. Although many methods have been reported, it remains a challenge for the assembly of designer-based TALE repeats in a fast, precise and cost-effective manner. We present an ULtiMATE (USER-based Ligation Mediated Assembly of TAL Effector) system for speedy and accurate assembly of customized TALE constructs. This method takes advantage of uracil-specific excision reagent (USER) to create multiple distinct sticky ends between any neighboring DNA fragments for specific ligation. With pre-assembled templates, multiple TALE DNA-binding domains could be efficiently assembled in order within hours with minimal manual operation. This system has been demonstrated to produce both functional TALENs for effective gene knockout and TALE-mediated gene-specific transcription activation (TALE-TA). The feature of both ease-of-operation and high efficiency of ULtiMATE system makes it not only an ideal method for biologic labs, but also an approach well suited for large-scale assembly of TALENs and any other TALE-based constructions.http://europepmc.org/articles/PMC3815405?pdf=render |
| spellingShingle | Junjiao Yang Pengfei Yuan Dingqiao Wen Ying Sheng Shiyou Zhu Yuezhou Yu Xiang Gao Wensheng Wei ULtiMATE system for rapid assembly of customized TAL effectors. PLoS ONE |
| title | ULtiMATE system for rapid assembly of customized TAL effectors. |
| title_full | ULtiMATE system for rapid assembly of customized TAL effectors. |
| title_fullStr | ULtiMATE system for rapid assembly of customized TAL effectors. |
| title_full_unstemmed | ULtiMATE system for rapid assembly of customized TAL effectors. |
| title_short | ULtiMATE system for rapid assembly of customized TAL effectors. |
| title_sort | ultimate system for rapid assembly of customized tal effectors |
| url | http://europepmc.org/articles/PMC3815405?pdf=render |
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