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Nat Neurosci. 2016 Mar;19(3):517-22. doi: 10.1038/nn.4235. Epub 2016 Feb 1.

Engineering microdeletions and microduplications by targeting segmental duplications with CRISPR.

Tai DJ1,2,3,4, Ragavendran A1,3,4, Manavalan P1,3,4, Stortchevoi A1,3,4, Seabra CM1,2,3,4,5, Erdin S1,3,4, Collins RL1,3,4, Blumenthal I1, Chen X6, Shen Y7, Sahin M8, Zhang C9, Lee C9,10, Gusella JF1,11,12, Talkowski ME1,2,3,4,11.

Author information

1
Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.
2
Department of Neurology, Harvard Medical School, Boston, Massachusetts, USA.
3
Psychiatric and Neurodevelopmental Genetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.
4
Department of Psychiatry, Massachusetts General Hospital, Boston, Massachusetts, USA.
5
Institute of Biomedical Sciences Abel Salazar, University of Porto, Porto, Portugal.
6
Beijing Municipal Key Laboratory of Child Development and Nutriomics, Capital Institute of Pediatrics, Beijing, China.
7
Department of Laboratory Medicine, Boston Children's Hospital, Boston, Massachusetts, USA.
8
Department of Neurology, Boston Children's Hospital, Boston, Massachusetts, USA.
9
The Jackson Laboratory for Genomic Medicine, Farmington, Connecticut, USA.
10
Department of Graduate Studies - Life Sciences, Ewha Womans University, Seoul, Korea.
11
Program in Medical and Population Genetics, Broad Institute of Harvard and MIT, Cambridge, Massachusetts, USA.
12
Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.

Abstract

Recurrent, reciprocal genomic disorders resulting from non-allelic homologous recombination (NAHR) between near-identical segmental duplications (SDs) are a major cause of human disease, often producing phenotypically distinct syndromes. The genomic architecture of flanking SDs presents a challenge for modeling these syndromes; however, the capability to efficiently generate reciprocal copy number variants (CNVs) that mimic NAHR would represent a valuable modeling tool. We describe here a CRISPR/Cas9 genome engineering method, single-guide CRISPR/Cas targeting of repetitive elements (SCORE), to model reciprocal genomic disorders and demonstrate its capabilities by generating reciprocal CNVs of 16p11.2 and 15q13.3, including alteration of one copy-equivalent of the SDs that mediate NAHR in vivo. The method is reproducible, and RNA sequencing reliably clusters transcriptional signatures from human subjects with in vivo CNVs and their corresponding in vitro models. This new approach will provide broad applicability for the study of genomic disorders and, with further development, may also permit efficient correction of these defects.

PMID:
26829649
PMCID:
PMC4903018
DOI:
10.1038/nn.4235
[Indexed for MEDLINE]
Free PMC Article

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