Open AccessTargeted molecular trait stacking in cotton through targeted double‐strand break induction
Author(s) -
D'Halluin Kathleen,
Vanderstraeten Chantal,
Hulle Jolien,
Rosolowska Joanna,
Den Brande Ilse,
Pennewaert Anouk,
D'Hont Kristel,
Bossut Martine,
Jantz Derek,
Ruiter Rene,
Broadhvest Jean
Publication year - 2013
Publication title -
plant biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.525
H-Index - 115
eISSN - 1467-7652
pISSN - 1467-7644
DOI - 10.1111/pbi.12085
Subject(s) - biology , genetics , crispr , genome editing , homologous recombination , genome , gene , locus (genetics) , dna , genome engineering , gene targeting , transcription activator like effector nuclease , computational biology
Summary 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.