A versatile reporter system for CRISPR-mediated chromosomal rearrangements

Yingxiang Li; Angela I. Park; Haiwei Mou; Cansu Colpan; Aizhan Bizhanova; Elliot Akama-Garren; Nik Joshi; Eric A. Hendrickson; David Feldser; Hao Yin; Daniel G. Anderson; Tyler Jacks; Zhiping Weng; Wen Xue

doi:10.1186/s13059-015-0680-7

A versatile reporter system for CRISPR-mediated chromosomal rearrangements

Yingxiang Li, Angela I. Park, Haiwei Mou, Cansu Colpan, Aizhan Bizhanova, Elliot Akama-Garren, Nik Joshi, Eric A. Hendrickson, David Feldser, Hao Yin, Daniel G. Anderson, Tyler Jacks, Zhiping Weng, Wen Xue

Biochemistry, Molecular Biology, and Biophysics (TMED)

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48 Scopus citations

Abstract

Although chromosomal deletions and inversions are important in cancer, conventional methods for detecting DNA rearrangements require laborious indirect assays. Here we develop fluorescent reporters to rapidly quantify CRISPR/Cas9-mediated deletions and inversions. We find that inversion depends on the non-homologous end-joining enzyme LIG4. We also engineer deletions and inversions for a 50 kb Pten genomic region in mouse liver. We discover diverse yet sequence-specific indels at the rearrangement fusion sites. Moreover, we detect Cas9 cleavage at the fourth nucleotide on the non-complementary strand, leading to staggered instead of blunt DNA breaks. These reporters allow mechanisms of chromosomal rearrangements to be investigated.

Original language	English (US)
Article number	111
Journal	Genome biology
Volume	16
Issue number	1
DOIs	https://doi.org/10.1186/s13059-015-0680-7
State	Published - May 28 2015

Bibliographical note

Publisher Copyright:
© 2015 Li et al.

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AU - Akama-Garren, Elliot

AU - Joshi, Nik

AU - Hendrickson, Eric A.

AU - Feldser, David

AU - Yin, Hao

AU - Anderson, Daniel G.

AU - Jacks, Tyler

AU - Weng, Zhiping

AU - Xue, Wen

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A versatile reporter system for CRISPR-mediated chromosomal rearrangements

Abstract

Bibliographical note

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