Multiplexed Targeted Genome Engineering Using a Universal Nuclease-Assisted Vector Integration System

Alexander Brown, Wendy S. Woods, Pablo Perez-Pinera

Research output: Contribution to journalArticle

Abstract

Engineered nucleases are capable of efficiently modifying complex genomes through introduction of targeted double-strand breaks. However, mammalian genome engineering remains limited by low efficiency of heterologous DNA integration at target sites, which is typically performed through homologous recombination, a complex, ineffective and costly process. In this study, we developed a multiplexable and universal nuclease-assisted vector integration system for rapid generation of gene knock outs using selection that does not require customized targeting vectors, thereby minimizing the cost and time frame needed for gene editing. Importantly, this system is capable of remodeling native mammalian genomes through integration of DNA, up to 50 kb, enabling rapid generation and screening of multigene knockouts from a single transfection. These results support that nuclease assisted vector integration is a robust tool for genome-scale gene editing that will facilitate diverse applications in synthetic biology and gene therapy.

Original languageEnglish (US)
Pages (from-to)582-588
Number of pages7
JournalACS synthetic biology
Volume5
Issue number7
DOIs
StatePublished - Jul 15 2016

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Keywords

  • CRISPR
  • DNA recombination
  • TALEN
  • gene editing
  • genome engineering
  • synthetic biology
  • targeted genome integration

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biochemistry, Genetics and Molecular Biology (miscellaneous)

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