BioTechniques, 2014-05, Vol.56 (5), p.263-268
2014
Details
- Autor(en) / Beteiligte
- Titel
- Efficient construction of rAAV-based gene targeting vectors by Golden Gate cloning
- Ist Teil von
- BioTechniques, 2014-05, Vol.56 (5), p.263-268
- Ort / Verlag
- England: Future Science Ltd
- Erscheinungsjahr
- 2014
- Link zum Volltext
- Quelle
- Taylor & Francis Journals Auto-Holdings Collection
- Beschreibungen/Notizen
- 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.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 0736-6205eISSN: 1940-9818DOI: 10.2144/000114169Titel-ID: cdi_proquest_miscellaneous_1654681747
- Format
- –
- Schlagworte
- Adeno-associated virus, Cinnamates - pharmacology, Cloning, Molecular - methods, Dependovirus - genetics, Drug Resistance, Microbial - genetics, Gene Knockout Techniques, gene targeting, Gene Targeting - methods, Genetic Engineering - methods, Genetic Vectors, Golden Gate cloning, Humans, Hygromycin B - analogs & derivatives, Hygromycin B - pharmacology, Neomycin - pharmacology, p53, rAAV, Reproducibility of Results, tau, tau Proteins - genetics