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Nat Methods. 2014 Jul;11(7):723-6. doi: 10.1038/nmeth.2969. Epub 2014 May 5.

CRISPR transcriptional repression devices and layered circuits in mammalian cells.

Author information

1
1] Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. [2] Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
2
Raytheon BBN Technologies, Cambridge, Massachusetts, USA.
3
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
4
1] Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. [2] Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. [3] Institute for the BioCentury, Korea Advanced Institute of Science and Technology, Daejeon, South Korea.
5
1] Bioinformatics Division, Center for Synthetic & Systems Biology, Tsinghua National Laboratory for Information Science and Technology, Beijing, China. [2] Department of Automation, Tsinghua University, Beijing, China.
6
Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
7
1] Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. [2] Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. [3] Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

Abstract

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.

PMID:
24797424
PMCID:
PMC4228775
DOI:
10.1038/nmeth.2969
[Indexed for MEDLINE]
Free PMC Article
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