Modularized CRISPR/dCas9 Effector Toolkit for Target-Specific Gene Regulation
Michael Agne(University of Freiburg), Ilona Blank(University of Freiburg), Alica Joana Emhardt(University of Freiburg), Christoph G. Gäbelein(University of Freiburg), Fenja Gawlas(University of Freiburg), Nadine Gillich(University of Freiburg), Patrick Gonschorek(University of Freiburg), Thomas J. Juretschke(University of Freiburg), Stefan Krämer(University of Freiburg), Natalie Louis(University of Freiburg), Anne Müller(University of Freiburg), Alina Rudorf(University of Freiburg), Lisa M. Schäfer(University of Freiburg), Manuel C. Scheidmann(University of Freiburg), Lisa J. Schmunk(University of Freiburg), Philipp Schwenk(University of Freiburg), Maximilian R. Stammnitz(University of Freiburg), Philipp Warmer(University of Freiburg), Wilfried Weber(University of Freiburg), Adrian Fischer(University of Freiburg), Beate Kaufmann(University of Freiburg), Hanna J. Wagner(University of Freiburg), Gerald Radziwill(University of Freiburg)
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Abstract
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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