title: Generation of chromosomal deletions in dicotyledonous plants employing a user-friendly genome editing toolkit.
abstract: Genome editing facilitated by Cas9-based RNA-guided nucleases (RGNs) is becoming an increasingly important and popular technique for reverse genetics in both model and non-model species. So far, RGNs were mainly applied for the induction of point mutations, and one major challenge consists in the detection of genome-edited individuals from a mutagenized population. Also, point mutations are not appropriate for functional dissection of non-coding DNA. Here, the multiplexing capacity of a newly developed genome editing toolkit was exploited for the induction of inheritable chromosomal deletions at six different loci in Nicotiana benthamiana and Arabidopsis. In both species, the preferential formation of small deletions was observed, suggesting reduced efficiency with increasing deletion size. Importantly, small deletions (<100 bp) were detected at high frequencies in N. benthamiana T0and Arabidopsis T2populations. Thus, targeting of small deletions by paired nucleases represents a simple approach for the generation of mutant alleles segregating as size polymorphisms in subsequent generations. Phenotypically selected deletions of up to 120 kb occurred at low frequencies in Arabidopsis, suggesting larger population sizes for the discovery of valuable alleles from addressing gene clusters or non-coding DNA for deletion by programmable nucleases.
authors: Department of Genetics, Martin Luther University Halle (Saale), Weinbergweg 10, 06120, Halle, Germany.
authors: Jana
authors: J
authors: Ordon
authors: Department of Genetics, Martin Luther University Halle (Saale), Weinbergweg 10, 06120, Halle, Germany.
authors: Johannes
authors: J
authors: Gantner
authors: Department of Genetics, Martin Luther University Halle (Saale), Weinbergweg 10, 06120, Halle, Germany.
authors: Jan
authors: J
authors: Kemna
authors: Department of Genetics, Martin Luther University Halle (Saale), Weinbergweg 10, 06120, Halle, Germany.
authors: Lennart
authors: L
authors: Schwalgun
authors: Department of Genetics, Martin Luther University Halle (Saale), Weinbergweg 10, 06120, Halle, Germany.
authors: Maik
authors: M
authors: Reschke
authors: Department of Genetics, Martin Luther University Halle (Saale), Weinbergweg 10, 06120, Halle, Germany.
authors: Jana
authors: J
authors: Streubel
authors: Department of Genetics, Martin Luther University Halle (Saale), Weinbergweg 10, 06120, Halle, Germany.
authors: Jens
authors: J
authors: Boch
authors: Department of Genetics, Martin Luther University Halle (Saale), Weinbergweg 10, 06120, Halle, Germany.
authors: Johannes
authors: J
authors: Stuttmann
mesh_terms: D000483:Alleles; D017360:Arabidopsis / Q000235:genetics*; D001483:Base Sequence; D064113:CRISPR-Cas Systems; D002869:Chromosome Aberrations; D032461:Chromosomes, Plant / Q000235:genetics*; D000072669:Gene Editing / Q000379:methods*; D018745:Genome, Plant / Q000235:genetics; D008957:Models, Genetic; D009154:Mutation; D010641:Phenotype; D030821:Plants, Genetically Modified; D011110:Polymorphism, Genetic; D014026:Nicotiana / Q000235:genetics*
keywords: 
  Arabidopsis thaliana
  ; 
  Nicotiana benthamiana
  ; CRISPR/Cas; EDS1; chromosomal deletion; plant immunity; technical advance