A key obstacle to creating sophisticated genetic circuits has been the lack of scalable device libraries. Here we present a modular transcriptional repression architecture based on clustered regularly interspaced palindromic repeats (CRISPR) system and examine approaches for regulated expression of guide RNAs in human cells. Subsequently we demonstrate that CRISPR regulatory devices can be layered to create functional cascaded circuits, which provide a valuable toolbox for engineering purposes.
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This work was supported by US National Institutes of Health grants 5R01CA155320-04 and P50 GM098792. We thank L. Wrobleska and P. Guye (Massachusetts Institute of Technology) for providing the initial intronic miRNA–based plasmid and the primary Cas9 construct, and for helpful discussions, and M. Graziano (Massachusetts Institute of Technology) for providing us the HEK293 cell lines that constitutively express rtTA3. J.H. was partially supported by the Intelligent Synthetic Biology Center of Global Frontier Project (2013M3A6A8073557) funded by the Ministry of Science, Information and Communication Technology and Future Planning of Korea.
The authors declare no competing financial interests.
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Kiani, S., Beal, J., Ebrahimkhani, M. et al. CRISPR transcriptional repression devices and layered circuits in mammalian cells. Nat Methods 11, 723–726 (2014). https://doi.org/10.1038/nmeth.2969
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