A versatile reporter system for CRISPR-mediated chromosomal rearrangements
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13059_2015_Article_680.pdf
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Author(s)
Li, Yingxiang
Mou, Haiwei
Colpan, Cansu
Bizhanova, Aizhan
Akama-Garren, Elliot
Joshi, Nik
Feldser, David M.
Yin, Hao
Weng, Zhiping
Xue, Wen
Date Issued
May 2015
Journal
Genome Biology
Publisher
BioMed Central
Citation
Li, Yingxiang, Angela I. Park, Haiwei Mou, Cansu Colpan, Aizhan Bizhanova, Elliot Akama-Garren, Nik Joshi, et al. “A Versatile Reporter System for CRISPR-Mediated Chromosomal Rearrangements.” Genome Biology 16, no. 1 (May 28, 2015).
Version
Final published version
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.
MIT Department
Massachusetts Institute of Technology. Institute for Medical Engineering & Science
Harvard University--MIT Division of Health Sciences and Technology
Massachusetts Institute of Technology. Department of Chemical Engineering
Koch Institute for Integrative Cancer Research at MIT
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1186/s13059-015-0680-7
