title: Highly efficient targeted chromosome deletions using CRISPR/Cas9.
abstract: 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 1 Mbp 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.
authors.affiliation: State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou, P. R. China; The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush Campus, Edinburgh, UK.
authors.forename: Zuyong
authors.initials: Z
authors.lastname: He
authors.forename: Chris
authors.initials: C
authors.lastname: Proudfoot
authors.forename: Alan J
authors.initials: AJ
authors.lastname: Mileham
authors.forename: David G
authors.initials: DG
authors.lastname: McLaren
authors.forename: C Bruce A
authors.initials: CB
authors.lastname: Whitelaw
authors.forename: Simon G
authors.initials: SG
authors.lastname: Lillico
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D002872:Chromosome Deletion; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D004247:DNA / Q000235:genetics*; D053903:DNA Breaks, Double-Stranded; D004720:Endonucleases / Q000235:genetics; D018390:Gene Targeting / Q000379:methods*; D005818:Genetic Engineering / Q000379:methods; D006801:Humans; D008969:Molecular Sequence Data; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics*
chemical_list: D000094704:RNA, Guide, CRISPR-Cas Systems; D004247:DNA; D004720:Endonucleases
keywords: CRISPR/Cas9; HPRT; large genomic deletion