Manipulation of gene expression on a genome-wide level is one of the most important systematic tools in the post-genome era. Such manipulations have largely been enabled by expression cloning approaches using sequence-verified cDNA libraries, large-scale RNA interference libraries (shRNA or siRNA) and zinc finger nuclease technologies. More recently, the CRISPR (clustered regularly interspaced short palindromic repeats) and CRISPR-associated (Cas)9-mediated gene editing technology has been described that holds great promise for future use of this technology in genomic manipulation. It was suggested that the CRISPR system has the potential to be used in high-throughput, large-scale loss of function screening. Here we discuss some of the challenges in engineering of CRISPR/Cas genomic libraries and some of the aspects that need to be addressed in order to use this technology on a high-throughput scale.
Keywords: CRISPR, Cas9, RNAi, gene silencing, gene editing, knockdown, screen, high-throughput
Citation: Heintze J, Luft C and Ketteler R (2013) A CRISPR CASe for high-throughput silencing. Front. Genet. 4:193. doi: 10.3389/fgene.2013.00193
Received: 13 June 2013; Accepted: 12 September 2013;
Published online: 07 October 2013.
Edited by:Rafael E. Carazo Salas, University of Cambridge, UK
Reviewed by:Neil Hukriede, University of Pittsburgh, USA
Copyright © 2013 Heintze, Luft and Ketteler. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: Robin Ketteler, Medical Research Council Laboratory for Molecular Cell Biology, University College London, Gower Street, London WC1E 6BT, UK e-mail: firstname.lastname@example.org