Multiplex Genome Engineering in Human Cells Using All-In-One CRISPR/Cas9 Vector System

Sci Rep. 2014 Jun 23;4:5400. doi: 10.1038/srep05400.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
  • Genetic Engineering / methods*
  • Genetic Vectors / genetics*
  • Genome, Human / genetics*
  • Humans
  • Transfection / methods*