Abstract
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.
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Acknowledgment
This work was supported by BBSRC UK mammalian Synthetic biology centre (BB/M018040/1). S.P. is a CRUK Senior Research Fellow (A17368).
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Gogolok, S., Köber, U., Pollard, S.M. (2018). STAR: A Simple TAL Effector Assembly Reaction Using Isothermal Assembly. In: Braman, J. (eds) Synthetic Biology. Methods in Molecular Biology, vol 1772. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7795-6_29
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DOI: https://doi.org/10.1007/978-1-4939-7795-6_29
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