Format

Send to

Choose Destination
See comment in PubMed Commons below
Nucleic Acids Res. 2013 Nov;41(21):9967-75. doi: 10.1093/nar/gkt758. Epub 2013 Aug 27.

Two- and three-input TALE-based AND logic computation in embryonic stem cells.

Author information

  • 1Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA, Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138, USA and Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA.

Abstract

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.

PMID:
23982518
PMCID:
PMC3834826
DOI:
10.1093/nar/gkt758
[PubMed - indexed for MEDLINE]
Free PMC Article
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Silverchair Information Systems Icon for PubMed Central
    Loading ...
    Support Center