Article
Highly efficient multiplex editing: one-shot generation of 8× Nicotiana benthamiana and 12× Arabidopsis mutants.
The Plant journal : for cell and molecular biology - 1 Apr 2021
Stuttmann Johannes, Barthel Karen, Martin Patrick, Ordon Jana, Erickson Jessica L, Herr Rosalie, Ferik Filiz, Kretschmer Carola, Berner Thomas, Keilwagen Jens, Marillonnet Sylvestre, Bonas Ulla
Abstract excerpt
Genome editing by RNA-guided nucleases, such as SpCas9, has been used in numerous different plant species. However, to what extent multiple independent loci can be targeted simultaneously by multiplexing has not been well documented. Here, we developed a toolkit, based on a highly intron-optimized zCas9i gene, which allows assembly of nuclease constructs expressing up to 32 single guide RNAs (sgRNAs). We used...
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