title: Engineering microdeletions and microduplications by targeting segmental duplications with CRISPR.
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.
authors.affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.|Department of Neurology, Harvard Medical School, Boston, Massachusetts, USA.|Psychiatric and Neurodevelopmental Genetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.|Department of Psychiatry, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Derek J C
authors.initials: DJ
authors.lastname: Tai
authors.affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.|Psychiatric and Neurodevelopmental Genetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.|Department of Psychiatry, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Ashok
authors.initials: A
authors.lastname: Ragavendran
authors.affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.|Psychiatric and Neurodevelopmental Genetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.|Department of Psychiatry, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Poornima
authors.initials: P
authors.lastname: Manavalan
authors.affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.|Psychiatric and Neurodevelopmental Genetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.|Department of Psychiatry, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Alexei
authors.initials: A
authors.lastname: Stortchevoi
authors.affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.|Department of Neurology, Harvard Medical School, Boston, Massachusetts, USA.|Psychiatric and Neurodevelopmental Genetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.|Department of Psychiatry, Massachusetts General Hospital, Boston, Massachusetts, USA.|Institute of Biomedical Sciences Abel Salazar, University of Porto, Porto, Portugal.
authors.forename: Catarina M
authors.initials: CM
authors.lastname: Seabra
authors.affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.|Psychiatric and Neurodevelopmental Genetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.|Department of Psychiatry, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Serkan
authors.initials: S
authors.lastname: Erdin
authors.affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.|Psychiatric and Neurodevelopmental Genetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.|Department of Psychiatry, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Ryan L
authors.identifier: http://orcid.org/0000-0003-1268-9995
authors.initials: RL
authors.lastname: Collins
authors.affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Ian
authors.initials: I
authors.lastname: Blumenthal
authors.affiliation: Beijing Municipal Key Laboratory of Child Development and Nutriomics, Capital Institute of Pediatrics, Beijing, China.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Chen
authors.affiliation: Department of Laboratory Medicine, Boston Children's Hospital, Boston, Massachusetts, USA.
authors.forename: Yiping
authors.initials: Y
authors.lastname: Shen
authors.affiliation: Department of Neurology, Boston Children's Hospital, Boston, Massachusetts, USA.
authors.forename: Mustafa
authors.identifier: http://orcid.org/0000-0001-7044-2953
authors.initials: M
authors.lastname: Sahin
authors.affiliation: The Jackson Laboratory for Genomic Medicine, Farmington, Connecticut, USA.
authors.forename: Chengsheng
authors.initials: C
authors.lastname: Zhang
authors.affiliation: The Jackson Laboratory for Genomic Medicine, Farmington, Connecticut, USA.|Department of Graduate Studies - Life Sciences, Ewha Womans University, Seoul, Korea.
authors.forename: Charles
authors.initials: C
authors.lastname: Lee
authors.affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.|Program in Medical and Population Genetics, Broad Institute of Harvard and MIT, Cambridge, Massachusetts, USA.|Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: James F
authors.initials: JF
authors.lastname: Gusella
authors.affiliation: Molecular Neurogenetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.|Department of Neurology, Harvard Medical School, Boston, Massachusetts, USA.|Psychiatric and Neurodevelopmental Genetics Unit, Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts, USA.|Department of Psychiatry, Massachusetts General Hospital, Boston, Massachusetts, USA.|Program in Medical and Population Genetics, Broad Institute of Harvard and MIT, Cambridge, Massachusetts, USA.
authors.forename: Michael E
authors.initials: ME
authors.lastname: Talkowski
mesh_terms: D001321:Autistic Disorder / Q000235:genetics*; D064113:CRISPR-Cas Systems / Q000235:genetics*; D002872:Chromosome Deletion; D025063:Chromosome Disorders / Q000235:genetics*; D002884:Chromosomes, Human, Pair 15 / Q000235:genetics; D002885:Chromosomes, Human, Pair 16 / Q000235:genetics; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics; D056915:DNA Copy Number Variations / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D006801:Humans; D008607:Intellectual Disability / Q000235:genetics*; D056916:Segmental Duplications, Genomic / Q000235:genetics*; D012640:Seizures / Q000235:genetics*; D017384:Sequence Deletion / Q000235:genetics*
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