Repositorium
Generation of chromosomal deletions in dicotyledonous plants employing a user-friendly genome editing toolkit
161
Journal Article / 2017
Ordon, Jana; Gantner, Johannes; Kemna, Jan; Schwalgun, Lennart; Reschke, Maik; Streubel, Jana; Boch, Jens; Stuttmann, Johannes
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 T0 and Arabidopsis T2 populations. 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.
Techniques
ID | Corresponding Author Country |
Plant Species | GE Technique Sequence Identifier |
Trait Type of Alteration |
Progress in Research Key Topic |
---|---|---|---|---|---|
343 |
Stuttmann, Johannes Germany |
Arabidopsis thaliana |
CRISPR/Cas9 DM2h |
No information SDN1 |
Basic research Basic research |
344 |
Stuttmann, Johannes Germany |
Arabidopsis thaliana |
CRISPR/Cas9 DMC2c |
No information SDN1 |
Basic research Basic research |
345 |
Stuttmann, Johannes Germany |
Arabidopsis thaliana |
CRISPR/Cas9 GUS |
No information SDN1 |
Basic research Basic research |
346 |
Stuttmann, Johannes Germany |
Nicotiana benthamiana |
CRISPR/Cas9 EDS1a/b |
No information SDN1 |
Basic research Basic research |
347 |
Stuttmann, Johannes Germany |
Nicotiana benthamiana |
CRISPR/Cas9 GUS |
GUS expression SDN1 |
Basic research Basic research |
348 |
Stuttmann, Johannes Germany |
Nicotiana benthamiana |
CRISPR/Cas9 PNbPAD4 |
No information SDN1 |
Basic research Basic research |