1. Modularized CRISPR/dCas9 effector toolkit for target-specific gene regulation.
- Author
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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, and Radziwill G
- Subjects
- HEK293 Cells, Humans, Plasmids genetics, Clustered Regularly Interspaced Short Palindromic Repeats genetics, Gene Expression Regulation genetics, Genetic Engineering methods
- 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.
- Published
- 2014
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