Engineering microdeletions and microduplications by targeting segmental duplications with CRISPR.

Document Type

Article

Publication Date

3-2016

JAX Source

Nat Neurosci 2016 Mar; 19(3):517-22.

Volume

19

Issue

3

First Page

517

Last Page

522

ISSN

1546-1726

PMID

26829649

Grant

CA034196, HG007497

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. Nat Neurosci 2016 Mar; 19(3):517-22.

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