| Autor: |
Gogolok S; MRC Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, UK., Köber U; MRC Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, UK., Pollard SM; MRC Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, UK. steven.pollard@ed.ac.uk. |
| Jazyk: |
angličtina |
| Zdroj: |
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2018; Vol. 1772, pp. 477-490. |
| DOI: |
10.1007/978-1-4939-7795-6_29 |
| Abstrakt: |
Transcription activator-like effectors (TALEs) contain programmable DNA-binding domains that can be fused to various effectors to manipulate genetic sequences or transcriptional state. However, the construction of plasmids encoding the modular DNA-binding domain remains challenging due to their repetitive nature. Here, we describe methods for a simple TALE assembly reaction (STAR) that uses a 68-part plasmid library to create TALEs binding to 17 bp target sequences. Manual production of many tens of TALEs can be achieved using a simple 8 h protocol, with full length sequence-verified plasmids available within a few days. This simple tale assembly reaction (STAR) provides a convenient method for generating tens to hundreds of TALENs or TALE-TFs without the need for large plasmid libraries or expensive liquid handling. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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