title: Inactivation of Cancer Mutations Utilizing CRISPR/Cas9.
abstract: Although whole-genome sequencing has uncovered a large number of mutations that drive tumorigenesis, functional ratification for most mutations remains sparse. Here, we present an approach to test functional relevance of tumor mutations employing CRISPR/Cas9. Combining comprehensive sgRNA design and an efficient reporter assay to nominate efficient and selective sgRNAs, we establish a pipeline to dissect roles of cancer mutations with potential applicability to personalized medicine and future therapeutic use.
authors: Affiliations of author: Medical Systems Biology, Medical Faculty Carl Gustav Carus, University cancer center, Technische Universitat Dresden, Dresden, Germany.
authors: Christina
authors: C
authors: Gebler
authors: Affiliations of author: Medical Systems Biology, Medical Faculty Carl Gustav Carus, University cancer center, Technische Universitat Dresden, Dresden, Germany.
authors: Tim
authors: T
authors: Lohoff
authors: Affiliations of author: Medical Systems Biology, Medical Faculty Carl Gustav Carus, University cancer center, Technische Universitat Dresden, Dresden, Germany.
authors: Maciej
authors: M
authors: Paszkowski-Rogacz
authors: Affiliations of author: Medical Systems Biology, Medical Faculty Carl Gustav Carus, University cancer center, Technische Universitat Dresden, Dresden, Germany.
authors: Jovan
authors: J
authors: Mircetic
authors: Affiliations of author: Medical Systems Biology, Medical Faculty Carl Gustav Carus, University cancer center, Technische Universitat Dresden, Dresden, Germany.
authors: Debojyoti
authors: D
authors: Chakraborty
authors: Affiliations of author: Medical Systems Biology, Medical Faculty Carl Gustav Carus, University cancer center, Technische Universitat Dresden, Dresden, Germany.
authors: Aylin
authors: A
authors: Camgoz
authors: Affiliations of author: Medical Systems Biology, Medical Faculty Carl Gustav Carus, University cancer center, Technische Universitat Dresden, Dresden, Germany.
authors: Martin V
authors: MV
authors: Hamann
authors: Affiliations of author: Medical Systems Biology, Medical Faculty Carl Gustav Carus, University cancer center, Technische Universitat Dresden, Dresden, Germany.
authors: Mirko
authors: M
authors: Theis
authors: Medical Faculty Carl Gustav Carus, Medizinische Klinik und Poliklinik I, Dresden, Germany.|German Cancer Research Center, Heidelberg and German Cancer Consortium partner site Dresden, Germany.
authors: Christian
authors: C
authors: Thiede
authors: Affiliations of author: Medical Systems Biology, Medical Faculty Carl Gustav Carus, University cancer center, Technische Universitat Dresden, Dresden, Germany.|Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany.|German Cancer Research Center, Heidelberg and German Cancer Consortium partner site Dresden, Germany.|National Center for Tumor Diseases, University Hospital Carl Gustav Carus, TU Dresden, Dresden, Germany.
authors: Frank
authors: F
authors: Buchholz
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D000076987:CRISPR-Associated Protein 9; D002277:Carcinoma / Q000235:genetics*; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D003110:Colonic Neoplasms / Q000235:genetics*; D019295:Computational Biology; D053837:DNA Cleavage; D004720:Endonucleases / Q000235:genetics / Q000378:metabolism; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000235:genetics*; D009154:Mutation / Q000235:genetics*; D009687:Nuclear Proteins / Q000235:genetics; D000090243:Nucleophosmin; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics*; D014162:Transfection
chemical_list: D001426:Bacterial Proteins; D009687:Nuclear Proteins; D000094704:RNA, Guide, CRISPR-Cas Systems; D000090243:Nucleophosmin; D049452:Green Fluorescent Proteins; C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf; D000076987:CRISPR-Associated Protein 9; C000606107:Cas9 endonuclease Streptococcus pyogenes; D004720:Endonucleases