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Here, we demonstrated 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 showed that targeting dCas9 to telomeres using sgTelo impeded DNA replication at telomeres and induced a pronounced increase of replication stress and DNA damage. Using Single-Molecule Telomere Assay via Optical Mapping (SMTA-OM), we investigated the genome-wide features of telomeres in the dCas9\/sgTelo cells and observed a dramatic increase of chromosome end fusions, including fusion\/ITS+ and fusion\/ITS-.Consistently, we also observed an increase in the formation of dicentric chromosomes, anaphase bridges, and intercellular telomeric chromosome bridges (ITCBs). Utilizing the dCas9\/sgTelo system, we uncovered many novel molecular and structural features of the ITCB and demonstrated 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.<\/jats:p>","DOI":"10.1101\/2024.04.03.587951","type":"posted-content","created":{"date-parts":[[2024,4,4]],"date-time":"2024-04-04T03:55:16Z","timestamp":1712202916000},"source":"Crossref","is-referenced-by-count":0,"title":["Elucidation of the molecular mechanism of the breakage-fusion-bridge (BFB) cycle using a CRISPR-dCas9 cellular model"],"prefix":"10.64898","author":[{"given":"Manrose","family":"Singh","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kaitlin","family":"Raseley","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexis M.","family":"Perez","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Danny","family":"MacKenzie","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Settapong T","family":"Kosiyatrakul","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sanket","family":"Desai","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Noelle","family":"Batista","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Navjot","family":"Guru","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Katherine K.","family":"Loomba","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Heba Z.","family":"Abid","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yilin","family":"Wang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lars","family":"Udo-Bellner","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Randy F.","family":"Stout","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Carl L.","family":"Schildkraut","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ming","family":"Xiao","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4515-2070","authenticated-orcid":false,"given":"Dong","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2024040609050525000_2024.04.03.587951v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(00)81683-9"},{"key":"2024040609050525000_2024.04.03.587951v1.2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2011.02.013"},{"key":"2024040609050525000_2024.04.03.587951v1.3","doi-asserted-by":"publisher","DOI":"10.1038\/25292"},{"key":"2024040609050525000_2024.04.03.587951v1.4","doi-asserted-by":"publisher","DOI":"10.1016\/j.gde.2022.101913"},{"issue":"1","key":"2024040609050525000_2024.04.03.587951v1.5","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1146\/annurev-cancerbio-070620-094029","article-title":"Cancer Genomic Rearrangements and Copy Number Alterations from Errors in Cell Division","volume":"6","year":"2022","journal-title":"Annual Review of Cancer Biology"},{"key":"2024040609050525000_2024.04.03.587951v1.6","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.090013497"},{"key":"2024040609050525000_2024.04.03.587951v1.7","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.25.8.405"},{"key":"2024040609050525000_2024.04.03.587951v1.8","doi-asserted-by":"publisher","DOI":"10.1093\/genetics\/26.2.234"},{"key":"2024040609050525000_2024.04.03.587951v1.9","doi-asserted-by":"crossref","unstructured":"Fernandez-Casanas, M. ; Chan, K. 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