A versatile reporter system for CRISPR-mediated chromosomal rearrangements

Genome Biology - Tập 16 - Trang 1-11 - 2015
Yingxiang Li1, Angela I. Park2, Haiwei Mou2, Cansu Colpan2, Aizhan Bizhanova2, Elliot Akama-Garren3, Nik Joshi3, Eric A. Hendrickson4, David Feldser5, Hao Yin3, Daniel G. Anderson3,6,7,8, Tyler Jacks3, Zhiping Weng1,9, Wen Xue2
1Department of Bioinformatics, School of Life Science and Technology, Tongji University, Shanghai, P. R. China
2RNA Therapeutics Institute and Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, USA
3David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, USA
4Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota Medical School, Minneapolis, USA
5Abramson Family Cancer Research Institute, University of Pennsylvania, Perelman School of Medicine, Philadelphia, USA
6Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, USA
7Harvard-MIT Division of Health Sciences & Technology, Cambridge, USA
8Institute of Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, USA
9Program in Bioinformatics and Integrative Biology, University of Massachusetts Medical School, Worcester, USA

Tóm tắt

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.

Tài liệu tham khảo

Thông tin
Thông tin xuất bản

Genome Biology

Tập 16

1-11

Thông tin tác giả