title: TALEN-Induced Translocations in Human Cells.
abstract: Induction of chromosomal translocations in human cells is of a great interest to study tumorigenesis and genome instability. Here, we explain in detail a method to induce translocations using the transcription activator-like effector nucleases (TALENs). We describe how to detect translocation formation by PCR, calculate translocation frequency by 96-well PCR screen, and analyze breakpoint junctions. When inducing cancer translocations, it is also possible to detect the fusion gene by FISH analysis or western blot.
authors.affiliation: Museum National d'Histoire Naturelle, 43 rue Cuvier, 75005, Paris, France.|CNRS, UMR7196, 43 rue Cuvier, 75005, Paris, France.|Inserm, U1154, 43 rue Cuvier, 75005, Paris, France.
authors.forename: Marion
authors.initials: M
authors.lastname: Piganeau
authors.affiliation: Museum National d'Histoire Naturelle, 43 rue Cuvier, 75005, Paris, France.|CNRS, UMR7196, 43 rue Cuvier, 75005, Paris, France.|Inserm, U1154, 43 rue Cuvier, 75005, Paris, France.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Renouf
authors.affiliation: Museum National d'Histoire Naturelle, 43 rue Cuvier, 75005, Paris, France.|CNRS, UMR7196, 43 rue Cuvier, 75005, Paris, France.|Inserm, U1154, 43 rue Cuvier, 75005, Paris, France.
authors.forename: Hind
authors.initials: H
authors.lastname: Ghezraoui
authors.affiliation: Museum National d'Histoire Naturelle, 43 rue Cuvier, 75005, Paris, France. ebrunet@mnhn.fr.|CNRS, UMR7196, 43 rue Cuvier, 75005, Paris, France. ebrunet@mnhn.fr.|Inserm, U1154, 43 rue Cuvier, 75005, Paris, France. ebrunet@mnhn.fr.
authors.forename: Erika
authors.initials: E
authors.lastname: Brunet
mesh_terms: D063646:Carcinogenesis / Q000235:genetics*; D056905:Chromosome Breakpoints; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics*; D004720:Endonucleases / Q000235:genetics; D018390:Gene Targeting / Q000379:methods; D042822:Genomic Instability / Q000235:genetics; D006801:Humans; D016297:Mutagenesis, Site-Directed; D015534:Trans-Activators / Q000737:chemistry / Q000235:genetics*; D014178:Translocation, Genetic / Q000235:genetics*
chemical_list: D004268:DNA-Binding Proteins; D015534:Trans-Activators; D004720:Endonucleases
keywords: 96-Well plate PCR screen; Breakpoint junction analysis; Fusion gene; T7-endonuclease assay; TALEN induced translocation