Engineering microdeletions and microduplications by targeting segmental duplications with CRISPR

Recurrent, reciprocal genomic disorders due to non-allelic homologous recombination (NAHR) are a major cause of human disease. The authors developed a CRISPR/Cas9 genome engineering method that directly targets segmental duplications and efficiently mimics the NAHR-mediated mechanism of microdeletio...

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Bibliographic Details
Published in:	Nature neuroscience Vol. 19; no. 3; pp. 517 - 522
Main Authors:	Tai, Derek J C, Ragavendran, Ashok, Manavalan, Poornima, Stortchevoi, Alexei, Seabra, Catarina M, Erdin, Serkan, Collins, Ryan L, Blumenthal, Ian, Chen, Xiaoli, Shen, Yiping, Sahin, Mustafa, Zhang, Chengsheng, Lee, Charles, Gusella, James F, Talkowski, Michael E
Format:	Journal Article
Language:	English
Published:	New York Nature Publishing Group US 01.03.2016 Nature Publishing Group
Subjects:	13/1 13/100 13/109 45/23 45/77 45/90 45/91 631/1647/1511 631/1647/767/722 631/61/338 Animal Genetics and Genomics Autistic Disorder - genetics Behavioral Sciences Biological Techniques Biomedicine Chromosome Deletion Chromosome Disorders - genetics Chromosomes, Human, Pair 15 - genetics Chromosomes, Human, Pair 16 - genetics Clustered Regularly Interspaced Short Palindromic Repeats - genetics Copy number variations CRISPR CRISPR-Cas Systems - genetics DNA Copy Number Variations - genetics Engineering Genetic engineering Genetic Engineering - methods Genetic research Genetic screening Genomes Hospitals Humans Identification and classification Innovations Intellectual Disability - genetics Laboratories Methods Mutation Neurobiology Neurosciences Proteins Segmental Duplications, Genomic - genetics Seizures - genetics Sequence Deletion - genetics technical-report United States > US Massachusetts
ISSN:	1097-6256, 1546-1726, 1546-1726
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Summary:	Recurrent, reciprocal genomic disorders due to non-allelic homologous recombination (NAHR) are a major cause of human disease. The authors developed a CRISPR/Cas9 genome engineering method that directly targets segmental duplications and efficiently mimics the NAHR-mediated mechanism of microdeletion and microduplication that occurs in vivo using 16p11.2 and 15q13.3 as proof-of-principle models. 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.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23
ISSN:	1097-6256 1546-1726 1546-1726
DOI:	10.1038/nn.4235