title: Modularized CRISPR/dCas9 effector toolkit for target-specific gene regulation.
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.
authors.affiliation: iGEM Team Freiburg 2013, ‡BIOSS Centre for Biological Signalling Studies, University of Freiburg , Schänzlestrasse 18, 79104 Freiburg, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Agne
authors.forename: Ilona
authors.initials: I
authors.lastname: Blank
authors.forename: Alica J
authors.initials: AJ
authors.lastname: Emhardt
authors.forename: Christoph G
authors.initials: CG
authors.lastname: Gäbelein
authors.forename: Fenja
authors.initials: F
authors.lastname: Gawlas
authors.forename: Nadine
authors.initials: N
authors.lastname: Gillich
authors.forename: Patrick
authors.initials: P
authors.lastname: Gonschorek
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Juretschke
authors.forename: Stefan D
authors.initials: SD
authors.lastname: Krämer
authors.forename: Natalie
authors.initials: N
authors.lastname: Louis
authors.forename: Anne
authors.initials: A
authors.lastname: Müller
authors.forename: Alina
authors.initials: A
authors.lastname: Rudorf
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Schäfer
authors.forename: Manuel C
authors.initials: MC
authors.lastname: Scheidmann
authors.forename: Lisa J
authors.initials: LJ
authors.lastname: Schmunk
authors.forename: Philipp M
authors.initials: PM
authors.lastname: Schwenk
authors.forename: Maximilian R
authors.initials: MR
authors.lastname: Stammnitz
authors.forename: Philipp M
authors.initials: PM
authors.lastname: Warmer
authors.forename: Wilfried
authors.initials: W
authors.lastname: Weber
authors.forename: Adrian
authors.initials: A
authors.lastname: Fischer
authors.forename: Beate
authors.initials: B
authors.lastname: Kaufmann
authors.forename: Hanna J
authors.initials: HJ
authors.lastname: Wagner
authors.forename: Gerald
authors.initials: G
authors.lastname: Radziwill
mesh_terms: D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D005786:Gene Expression Regulation / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D010957:Plasmids / Q000235:genetics