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Modularized CRISPR/dCas9 effector toolkit for target-specific gene regulation

ACS Synth Biol. 2015; 
Agne M, Blank I, Emhardt AJ, Gäbelein CG, Gawlas F, Gillich N, Gonschorek P, Juretschke TJ, Krämer SD, Louis N, Müller A, Rudorf A, Schäfer LM, Scheidmann MC, Schmunk LJ, Schwenk PM, Stammnitz MR, Warmer PM, Weber W, Fischer A, Kaufmann B, Wagner HJ, Radziwill G.
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Gene Synthesis This work was generously supported by the Excellence Initiative of the German Federal and State Governments (EXC-294, BIOSSCentre for Biological Signalling Studies), the Faculty of Medicine/ University of Freiburg and the Verband der Freunde der Universitat Freiburg. Additional funding was granted by A. ̈ Hartenstein, Anders + Redelfs, Biozym, Corning, Eppendorf, ERA SynBio, Genaxxon Bioscience, Geneious, GenScript, Gilson, Greiner Bio-One, HPS Labor und Bü rositzmö bel, Kuhnle, Life Technologies, MathWorks, New England BioLabs, Peprotech, Promega, Roche, Roth, Scienova, Stiftungen Landesbank Baden-Wü rttemberg, Thermo Scientific, WesoMed and Zymo Research. Get A Quote

摘要

The ability to control mammalian genes in a synergistic mode using synthetic transcription factors is highly desirable in fields of tissue engineering, stem cell reprogramming and fundamental research. In this study, we developed a standardized toolkit utilizing an engineered CRISPR/Cas9 system that enables customizable gene regulation in mammalian cells. The RNA-guided dCas9 protein was implemented as a programmable transcriptional activator or repressor device, including targeting of endogenous loci. For facile assembly of single or multiple CRISPR RNAs, our toolkit comprises a modular RNAimer plasmid, which encodes the required noncoding RNA components.

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