Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector system

• DOI: 10.1038/srep05400
• Publication Date: 2014-06-23T09:18:00Z
• Authors: Tetsushi Sakuma; Ayami Nishikawa; Satoshi Kume; Kazuaki Chayama; Takashi Yamamoto
• Keywords: 0301 basic medicine; 03 medical and health sciences; Genome, Human; Genetic Vectors; Humans; Clustered Regularly Interspaced Short Palindromic Repeats; Genetic Engineering; Transfection; Article

CRISPR/Cas9-mediated genome editing is a next-generation strategy for genetic modifications, not only for single gene targeting, but also for multiple targeted mutagenesis. To make the most of the multiplexity of CRISPR/Cas9, we established a system for constructing all-in-one expression vectors containing multiple guide RNA expression cassettes and a Cas9 nuclease/nickase expression cassette. We further demonstrated successful examples of multiple targeting including chromosomal deletions in human cells using the all-in-one CRISPR/Cas9 vectors constructed with our novel system. Our system provides an efficient targeting strategy for multiplex genome/epigenome editing, simultaneous activation/repression of multiple genes, and beyond.