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

Research output: Contribution to journalArticle

31 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 languageEnglish (US)
Article number111
JournalGenome biology
Volume16
Issue number1
DOIs
StatePublished - May 28 2015

    Fingerprint

Cite this

Li, Y., Park, A. I., Mou, H., Colpan, C., Bizhanova, A., Akama-Garren, E., Joshi, N., Hendrickson, E. A., Feldser, D., Yin, H., Anderson, D. G., Jacks, T., Weng, Z., & Xue, W. (2015). A versatile reporter system for CRISPR-mediated chromosomal rearrangements. Genome biology, 16(1), [111]. https://doi.org/10.1186/s13059-015-0680-7