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

  • Sakuma T
  • Nishikawa A
  • Kume S
 et al. 
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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.

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Authors

  • Tetsushi Sakuma

  • Ayami Nishikawa

  • Satoshi Kume

  • Kazuaki Chayama

  • Takashi Yamamoto

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