Journal article
Elucidation of the molecular mechanism of the breakage-fusion-bridge (BFB) cycle using a CRISPR-dCas9 cellular model
Nucleic acids research, v 52(19), pp 11689-11703
28 Oct 2024
PMID: 39193906
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Chromosome instability (CIN) is frequently observed in many tumors. The breakage-fusion-bridge (BFB) cycle has been proposed to be one of the main drivers of CIN during tumorigenesis and tumor evolution. However, the detailed mechanism for the individual steps of the BFB cycle warrants further investigation. Here, we demonstrate that a nuclease-dead Cas9 (dCas9) coupled with a telomere-specific single-guide RNA (sgTelo) can be used to model the BFB cycle. First, we show that targeting dCas9 to telomeres using sgTelo impedes DNA replication at telomeres and induces a pronounced increase of replication stress and DNA damage. Using Single-Molecule Telomere Assay via Optical Mapping (SMTA-OM), we investigate the genome-wide features of telomeres in the dCas9/sgTelo cells and observe a dramatic increase of chromosome end fusions, including fusion/ITS+ and fusion/ITS-. Consistently, we also observe an increase in the formation of dicentric chromosomes, anaphase bridges, and intercellular telomeric chromosome bridges (ITCBs). Utilizing the dCas9/sgTelo system, we uncover many interesting molecular and structural features of the ITCB and demonstrate that multiple DNA repair pathways are implicated in the formation of ITCBs. Our studies shed new light on the molecular mechanisms of the BFB cycle, which will advance our understanding of tumorigenesis, tumor evolution, and drug resistance.
Metrics
1 Record Views
Details
- Title
- Elucidation of the molecular mechanism of the breakage-fusion-bridge (BFB) cycle using a CRISPR-dCas9 cellular model
- Creators
- Manrose Singh - New York Institute of TechnologyKaitlin Raseley - Drexel UniversityAlexis M Perez - New York Institute of TechnologyDanny MacKenzie - New York Institute of TechnologySettapong T Kosiyatrakul - Albert Einstein College of MedicineSanket Desai - New York Institute of TechnologyNoelle Batista - New York Institute of TechnologyNavjot Guru - New York Institute of TechnologyKatherine K Loomba - New York Institute of TechnologyHeba Z Abid - Drexel UniversityYilin Wang - Drexel UniversityLars Udo-Bellner - New York Institute of TechnologyRandy F Stout - New York Institute of TechnologyCarl L Schildkraut - Albert Einstein College of MedicineMing Xiao - Drexel UniversityDong Zhang - New York Institute of Technology
- Publication Details
- Nucleic acids research, v 52(19), pp 11689-11703
- Publisher
- Oxford University Press
- Grant note
- New York Institute of Technology P30 CA013330 / NCI NIH HHS
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- School of Biomedical Engineering and Science
- Web of Science ID
- WOS:001300066500001
- Scopus ID
- 2-s2.0-85208160554
- Other Identifier
- 991022202499004721
UN Sustainable Development Goals (SDGs)
This publication has contributed to the advancement of the following goals:
Source: SDGs in the Output
InCites Highlights
Data related to this publication, from InCites Benchmarking & Analytics tool:
- Collaboration types
- Domestic collaboration
- Web of Science research areas
- Biochemistry & Molecular Biology