Journal
NUCLEIC ACIDS RESEARCH
Volume 41, Issue 21, Pages 9967-9975Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkt758
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Funding
- European Molecular Biology Organization Fellowship
- Human Frontier Science Program Fellowship
- National Science Foundation Graduate Research Fellowship
- Natural Sciences and Engineering Research Council of Canada Postdoctoral Fellowship
- National Institutes of Health [GM007598]
- NIH [R01GM036373]
- Defense Advanced Research Projects Agency (DARPA) [4500000572]
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Biological computing circuits can enhance our ability to control cellular functions and have potential applications in tissue engineering and medical treatments. Transcriptional activator-like effectors (TALEs) represent attractive components of synthetic gene regulatory circuits, as they can be designed de novo to target a given DNA sequence. We here demonstrate that TALEs can perform Boolean logic computation in mammalian cells. Using a split-intein protein-splicing strategy, we show that a functional TALE can be reconstituted from two inactive parts, thus generating two-input AND logic computation. We further demonstrate three-piece intein splicing in mammalian cells and use it to perform three-input AND computation. Using methods for random as well as targeted insertion of these relatively large genetic circuits, we show that TALE-based logic circuits are functional when integrated into the genome of mouse embryonic stem cells. Comparing construct variants in the same genomic context, we modulated the strength of the TALE-responsive promoter to improve the output of these circuits. Our work establishes split TALEs as a tool for building logic computation with the potential of controlling expression of endogenous genes or transgenes in response to a combination of cellular signals.
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