Highly efficient targeted chromosome deletions using CRISPR/Cas9

Zuyong He, Christopher Proudfoot, Alan J. Mileham, David G. Mclaren, C. Bruce A Whitelaw, Simon G. Lillico

Research output: Contribution to journalArticlepeer-review


The CRISPR/Cas9 system has emerged as an intriguing new technology for genome engineering. It utilizes the bacterial endonuclease Cas9 which, when delivered to eukaryotic cells in conjunction with a user-specified small guide RNA (gRNA), cleaves the chromosomal DNA at the target site. Here we show that concurrent delivery of gRNAs designed to target two different sites in a human chromosome introduce DNA double-strand breaks in the chromosome and give rise to targeted deletions of the intervening genomic segment. Predetermined genomic DNA segments ranging from several-hundred base pairs to 1Mbp can be precisely deleted at frequencies of 1-10%, with no apparent correlation between the size of the deleted fragment and the deletion frequency. The high efficiency of this technique holds promise for large genomic deletions that could be useful in generation of cell and animal models with engineered chromosomes.

Original languageEnglish
Pages (from-to)1060-1064
JournalBiotechnology and Bioengineering
Issue number5
Early online date23 Dec 2014
Publication statusPublished - 1 Jan 2015


  • CRISPR/Cas9
  • HPRT
  • Large genomic deletion

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