title: Targeted molecular trait stacking in cotton through targeted double-strand break induction.
abstract: Recent developments of tools for targeted genome modification have led to new concepts in how multiple traits can be combined. Targeted genome modification is based on the use of nucleases with tailor-made specificities to introduce a DNA double-strand break (DSB) at specific target loci. A re-engineered meganuclease was designed for specific cleavage of an endogenous target sequence adjacent to a transgenic insect control locus in cotton. The combination of targeted DNA cleavage and homologous recombination-mediated repair made precise targeted insertion of additional trait genes (hppd, epsps) feasible in cotton. Targeted insertion events were recovered at a frequency of about 2% of the independently transformed embryogenic callus lines. We further demonstrated that all trait genes were inherited as a single genetic unit, which will simplify future multiple-trait introgression.
authors.affiliation: Bayer CropScience N.V., Gent, Belgium.
authors.forename: Kathleen
authors.initials: K
authors.lastname: D'Halluin
authors.forename: Chantal
authors.initials: C
authors.lastname: Vanderstraeten
authors.forename: Jolien
authors.initials: J
authors.lastname: Van Hulle
authors.forename: Joanna
authors.initials: J
authors.lastname: Rosolowska
authors.forename: Ilse
authors.initials: I
authors.lastname: Van Den Brande
authors.forename: Anouk
authors.initials: A
authors.lastname: Pennewaert
authors.forename: Kristel
authors.initials: K
authors.lastname: D'Hont
authors.forename: Martine
authors.initials: M
authors.lastname: Bossut
authors.forename: Derek
authors.initials: D
authors.lastname: Jantz
authors.forename: Rene
authors.initials: R
authors.lastname: Ruiter
authors.forename: Jean
authors.initials: J
authors.lastname: Broadhvest
mesh_terms: D001483:Base Sequence; D053903:DNA Breaks, Double-Stranded; D005818:Genetic Engineering / Q000379:methods; D018745:Genome, Plant; D003368:Gossypium / Q000235:genetics*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010572:Pest Control, Biological; D030821:Plants, Genetically Modified; D059767:Recombinational DNA Repair; D017422:Sequence Analysis, DNA
keywords: cotton; gene targeting; homologous recombination; meganuclease; targeted DNA double-strand break induction and repair; targeted molecular trait stacking