Efficient construction of rAAV-based gene targeting vectors by Golden Gate cloning
The recombinant adeno-associated virus (rAAV) has proven to be an efficient and attractive tool for targeted genome engineering. Here we present a novel method employing the Golden Gate cloning strategy for fast and efficient construction of rAAV-based gene knockout or single-nucleotide knockin vect...
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| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2014-05-01
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| Series: | BioTechniques |
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| Online Access: | https://www.future-science.com/doi/10.2144/000114169 |
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| _version_ | 1850152490158260224 |
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| author | Yonglun Luo Lin Lin Lars Bolund Charlotte Brandt Sørensen |
| author_facet | Yonglun Luo Lin Lin Lars Bolund Charlotte Brandt Sørensen |
| author_sort | Yonglun Luo |
| collection | DOAJ |
| description | The recombinant adeno-associated virus (rAAV) has proven to be an efficient and attractive tool for targeted genome engineering. Here we present a novel method employing the Golden Gate cloning strategy for fast and efficient construction of rAAV-based gene knockout or single-nucleotide knockin vectors. Two vectors, pGolden-Neo and pGolden-Hyg, were generated as common assembling modules to confer antibiotic resistance to the targeting vector. To validate the method, we then generated two rAAV-based targeting vectors: pAAV-pTP53-KO and pAAV-hTauP301L-KI. Furthermore, we generated a pGolden-AAV plasmid that allows one-step generation of an rAAV-based targeting vector. Our new methodology for rAAV targeting vector assembly is efficient, accurate, time-saving, and cost-effective. |
| format | Article |
| id | doaj-art-c4d162103e1a4c6e99381c703455ad81 |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2014-05-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-c4d162103e1a4c6e99381c703455ad812025-08-20T02:25:58ZengTaylor & Francis GroupBioTechniques0736-62051940-98182014-05-0156526326810.2144/000114169Efficient construction of rAAV-based gene targeting vectors by Golden Gate cloningYonglun Luo0Lin Lin1Lars Bolund2Charlotte Brandt Sørensen31Department of Biomedicine, Faculty of Heath, Aarhus University, Aarhus, Denmark1Department of Biomedicine, Faculty of Heath, Aarhus University, Aarhus, Denmark1Department of Biomedicine, Faculty of Heath, Aarhus University, Aarhus, Denmark1Department of Biomedicine, Faculty of Heath, Aarhus University, Aarhus, DenmarkThe recombinant adeno-associated virus (rAAV) has proven to be an efficient and attractive tool for targeted genome engineering. Here we present a novel method employing the Golden Gate cloning strategy for fast and efficient construction of rAAV-based gene knockout or single-nucleotide knockin vectors. Two vectors, pGolden-Neo and pGolden-Hyg, were generated as common assembling modules to confer antibiotic resistance to the targeting vector. To validate the method, we then generated two rAAV-based targeting vectors: pAAV-pTP53-KO and pAAV-hTauP301L-KI. Furthermore, we generated a pGolden-AAV plasmid that allows one-step generation of an rAAV-based targeting vector. Our new methodology for rAAV targeting vector assembly is efficient, accurate, time-saving, and cost-effective.https://www.future-science.com/doi/10.2144/000114169gene targetingrAAVGolden Gate cloningp53tau |
| spellingShingle | Yonglun Luo Lin Lin Lars Bolund Charlotte Brandt Sørensen Efficient construction of rAAV-based gene targeting vectors by Golden Gate cloning BioTechniques gene targeting rAAV Golden Gate cloning p53 tau |
| title | Efficient construction of rAAV-based gene targeting vectors by Golden Gate cloning |
| title_full | Efficient construction of rAAV-based gene targeting vectors by Golden Gate cloning |
| title_fullStr | Efficient construction of rAAV-based gene targeting vectors by Golden Gate cloning |
| title_full_unstemmed | Efficient construction of rAAV-based gene targeting vectors by Golden Gate cloning |
| title_short | Efficient construction of rAAV-based gene targeting vectors by Golden Gate cloning |
| title_sort | efficient construction of raav based gene targeting vectors by golden gate cloning |
| topic | gene targeting rAAV Golden Gate cloning p53 tau |
| url | https://www.future-science.com/doi/10.2144/000114169 |
| work_keys_str_mv | AT yonglunluo efficientconstructionofraavbasedgenetargetingvectorsbygoldengatecloning AT linlin efficientconstructionofraavbasedgenetargetingvectorsbygoldengatecloning AT larsbolund efficientconstructionofraavbasedgenetargetingvectorsbygoldengatecloning AT charlottebrandtsørensen efficientconstructionofraavbasedgenetargetingvectorsbygoldengatecloning |