التفاصيل البيبلوغرافية
| العنوان: |
Generating new fanca-deficient hnscc cell lines by genomic editing recapitulates the cellular phenotypes of fanconi anemia |
| المؤلفون: |
Errazquin, Ricardo, Sieiro, Esther, Moreno Sánchez, Pilar, Ramírez de Haro, Ma. José, Lorz, Corina, Peral, Jorge, Ortiz, Jessica, Casado, José A., Roman-Rodriguez, Francisco J., Hanenberg, Helmut, Río, Paula, Surrallés i Calonge, Jordi, Segrelles, Carmen, Garcia-Escudero, Ramon, Universitat Autònoma de Barcelona |
| سنة النشر: |
2021 |
| المجموعة: |
Universitat Autònoma de Barcelona: Dipòsit Digital de Documents de la UAB |
| مصطلحات موضوعية: |
Fanconi anemia, CRISPR/Cas9, Gene editing, FANCA, Head and neck cancer |
| الوصف: |
Fanconi anemia (FA) patients have an exacerbated risk of head and neck squamous cell carcinoma (HNSCC). Treatment is challenging as FA patients display enhanced toxicity to standard treatments, including radio/chemotherapy. Therefore, better therapies as well as new disease models are urgently needed. We have used CRISPR/Cas9 editing tools in order to interrupt the human FANCA gene by the generation of insertions/deletions (indels) in exon 4 in two cancer cell lines from sporadic HNSCC having no mutation in FA-genes: CAL27 and CAL33 cells. Our approach allowed efficient editing, subsequent purification of single-cell clones, and Sanger sequencing validation at the edited locus. Clones having frameshift indels in homozygosis did not express FANCA protein and were selected for further analysis. When compared with parental CAL27 and CAL33, FANCA-mutant cell clones displayed a FA-phenotype as they (i) are highly sensitive to DNA interstrand crosslink (ICL) agents such as mitomycin C (MMC) or cisplatin(ii) do not monoubiquitinate FANCD2 upon MMC treatment and therefore (iii) do not form FANCD2 nuclear foci, and (iv) they display increased chromosome fragility and G2 arrest after diepoxybutane (DEB) treatment. These FANCA-mutant clones display similar growth rates as their parental cells. Interestingly, mutant cells acquire phenotypes associated with more aggressive disease, such as increased migration in wound healing assays. Therefore, CAL27 and CAL33 cells with FANCA mutations are phenocopies of FA-HNSCC cells. |
| نوع الوثيقة: |
article in journal/newspaper |
| وصف الملف: |
application/pdf |
| اللغة: |
English |
| تدمد: |
20734425 |
| Relation: |
Instituto de Salud Carlos III PI18/00263; Instituto de Salud Carlos III CB16/12/00228; Genes; Vol. 12 Núm. 4 (april 2021), p. 548; https://ddd.uab.cat/record/250110; urn:10.3390/genes12040548; urn:oai:ddd.uab.cat:250110; urn:scopus_id:85104576776; urn:articleid:20734425v12n4p548; urn:pmid:33918752; urn:pmc-uid:8069753; urn:pmcid:PMC8069753; urn:oai:pubmedcentral.nih.gov:8069753; urn:oai:egreta.uab.cat:publications/ece886b5-5cef-45e7-9beb-df956c8c3f7c |
| الاتاحة: |
https://ddd.uab.cat/record/250110 |
| Rights: |
open access ; Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original. ; https://creativecommons.org/licenses/by/4.0/ |
| رقم الانضمام: |
edsbas.108D0DFF |
| قاعدة البيانات: |
BASE |
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