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A versatile reporter system for CRISPR-mediated chromosomal rearrangements.

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.

SUBMITTER: Li Y

PROVIDER: S-EPMC4465146 | biostudies-literature | 2015 May

REPOSITORIES: biostudies-literature

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A versatile reporter system for CRISPR-mediated chromosomal rearrangements.

Li Yingxiang Y Park Angela I AI Mou Haiwei H Colpan Cansu C Bizhanova Aizhan A Akama-Garren Elliot E Joshi Nik N Hendrickson Eric A EA Feldser David D Yin Hao H Anderson Daniel G DG Jacks Tyler T Weng Zhiping Z Xue Wen W

Genome biology 20150528

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 rearrangeme ...[more]

PMID: 26018130

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