Deletions, Inversions, Duplications: Engineering of Structural Variants using CRISPR/Cas in Mice

被引：173

作者：

Kraft, Katerina ^{[1
]}

Geuer, Sinje ^{[1
,2
]}

Will, Anja J. ^{[1
,2
]}

Chan, Wing Lee ^{[2
]}

Paliou, Christina ^{[1
]}

Borschiwer, Marina ^{[1
]}

Harabula, Izabela ^{[1
]}

Wittler, Lars ^{[1
]}

Franke, Martin ^{[1
,2
]}

Ibrahim, Daniel M. ^{[1
,3
]}

Kragesteen, Bjort K. ^{[1
,2
,3
]}

Spielmann, Malte ^{[1
,2
,3
,4
]}

Mundlos, Stefan ^{[1
,2
,3
,4
]}

Lupianez, Dario G. ^{[1
]}

Andrey, Guillaume ^{[1
]}

机构：

[1] Max Planck Inst Mol Genet, D-14195 Berlin, Germany

[2] Charite, Inst Med & Human Genet, D-13353 Berlin, Germany

[3] Charite, Berlin Brandenburg Ctr Regenerat Therapies BCRT, D-13353 Berlin, Germany

[4] Charite, Berlin Brandenburg Sch Regenerat Therapies BSRT, D-13353 Berlin, Germany

来源：

CELL REPORTS | 2015年 / 10卷 / 05期

基金：

瑞士国家科学基金会;

关键词：

EMBRYONIC STEM-CELLS; GENOMIC REARRANGEMENTS; CHROMOSOMAL DELETIONS; REGULATORY LANDSCAPE; ENDONUCLEASE; GENERATION; ZEBRAFISH; ENHANCER; DISEASE; LOCUS;

D O I：

10.1016/j.celrep.2015.01.016

中图分类号：

Q2 [细胞生物学];

学科分类号：

071013 [干细胞生物学];

摘要：

Structural variations (SVs) contribute to the variability of our genome and are often associated with disease. Their study in model systems was hampered until now by labor-intensive genetic targeting procedures and multiple mouse crossing steps. Here we present the use of CRISPR/Cas for the fast (10 weeks) and efficient generation of SVs in mice. We specifically produced deletions, inversions, and also duplications at six different genomic loci ranging from 1.1 kb to 1.6 Mb with efficiencies up to 42%. After PCR-based selection, clones were successfully used to create mice via aggregation. To test the practicability of the method, we reproduced a human 500 kb disease- associated deletion and were able to recapitulate the human phenotype in mice. Furthermore, we evaluated the regulatory potential of a large genomic interval by deleting a 1.5 Mb fragment. The method presented permits rapid in vivo modeling of genomic rearrangements.

引用

页码：833 / 839

页数：7

共 24 条

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