title: Highly specific contractions of a single CAG/CTG trinucleotide repeat by TALEN in yeast.
abstract: Trinucleotide repeat expansions are responsible for more than two dozens severe neurological disorders in humans. A double-strand break between two short CAG/CTG trinucleotide repeats was formerly shown to induce a high frequency of repeat contractions in yeast. Here, using a dedicated TALEN, we show that induction of a double-strand break into a CAG/CTG trinucleotide repeat in heterozygous yeast diploid cells results in gene conversion of the repeat tract with near 100% efficacy, deleting the repeat tract. Induction of the same TALEN in homozygous yeast diploids leads to contractions of both repeats to a final length of 3-13 triplets, with 100% efficacy in cells that survived the double-strand breaks. Whole-genome sequencing of surviving yeast cells shows that the TALEN does not increase mutation rate. No other CAG/CTG repeat of the yeast genome showed any length alteration or mutation. No large genomic rearrangement such as aneuploidy, segmental duplication or translocation was detected. It is the first demonstration that induction of a TALEN in an eukaryotic cell leads to shortening of trinucleotide repeat tracts to lengths below pathological thresholds in humans, with 100% efficacy and very high specificity.
authors.affiliation: Institut Pasteur, Unité de Génétique Moléculaire des Levures, Département Génomes & Génétique, Paris, France; Sorbonne Universités, UPMC Univ Paris 6, IFD, Paris, France; CNRS, UMR3525, Paris, France.
authors.forename: Guy-Franck
authors.initials: GF
authors.lastname: Richard
authors.affiliation: Institut Pasteur, Unité de Génétique Moléculaire des Levures, Département Génomes & Génétique, Paris, France; Sorbonne Universités, UPMC Univ Paris 6, IFD, Paris, France; CNRS, UMR3525, Paris, France.
authors.forename: David
authors.initials: D
authors.lastname: Viterbo
authors.affiliation: Institut Pasteur, Unité de Génétique Moléculaire des Levures, Département Génomes & Génétique, Paris, France; Sorbonne Universités, UPMC Univ Paris 6, IFD, Paris, France; CNRS, UMR3525, Paris, France.
authors.forename: Varun
authors.initials: V
authors.lastname: Khanna
authors.affiliation: Institut Pasteur, Unité de Génétique Moléculaire des Levures, Département Génomes & Génétique, Paris, France; Sorbonne Universités, UPMC Univ Paris 6, IFD, Paris, France; CNRS, UMR3525, Paris, France.
authors.forename: Valentine
authors.initials: V
authors.lastname: Mosbach
authors.affiliation: Institut Pasteur, Unité de Génétique Moléculaire des Levures, Département Génomes & Génétique, Paris, France; Sorbonne Universités, UPMC Univ Paris 6, IFD, Paris, France; CNRS, UMR3525, Paris, France.
authors.forename: Lauriane
authors.initials: L
authors.lastname: Castelain
authors.affiliation: Institut Pasteur, Unité de Génétique Moléculaire des Levures, Département Génomes & Génétique, Paris, France; Sorbonne Universités, UPMC Univ Paris 6, IFD, Paris, France; CNRS, UMR3525, Paris, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Dujon
mesh_terms: D053903:DNA Breaks, Double-Stranded; D005656:Fungal Proteins / Q000378:metabolism*; D005838:Genotype; D059785:Karyotype; D059645:Mutation Rate; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D019680:Trinucleotide Repeat Expansion; D018911:Trinucleotide Repeats; D015003:Yeasts / Q000235:genetics* / Q000378:metabolism*
chemical_list: D005656:Fungal Proteins
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