A versatile reporter system for CRISPR-mediated chromosomal rearrangements
Li, Yingxiang ; Park, Angela I. ; Mou, Haiwei ; Colpan, Cansu ; Bizhanova, Aizhan ; Akama-Garren, Elliot ; Joshi, Nik ; Hendrickson, Eric A. ; Feldser, David ; Yin, Hao ... show 4 more
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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.
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Genome Biol. 2015 May 28;16:111. doi: 10.1186/s13059-015-0680-7. Link to article on publisher's site