Generation of chromosomal deletions in dicotyledonous plants employing a user-friendly genome editing toolkit

Research output: Contribution to journalArticleResearchpeer review

Authors

  • Jana Ordon
  • Johannes Gantner
  • Jan Kemna
  • Lennart Schwalgun
  • Maik Reschke
  • Jana Streubel
  • Jens Boch
  • Johannes Stuttmann

External Research Organisations

  • Martin Luther University Halle-Wittenberg
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Details

Original languageEnglish
Pages (from-to)155-168
Number of pages14
JournalPlant Journal
Volume89
Issue number1
Publication statusPublished - 14 Nov 2016
Externally publishedYes

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 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.

Keywords

    Arabidopsis thaliana, chromosomal deletion, CRISPR/Cas, EDS1, Nicotiana benthamiana, plant immunity, technical advance

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Genetics
  • Agricultural and Biological Sciences(all)
  • Plant Science
  • Biochemistry, Genetics and Molecular Biology(all)
  • Cell Biology

Cite this

Generation of chromosomal deletions in dicotyledonous plants employing a user-friendly genome editing toolkit. / Ordon, Jana; Gantner, Johannes; Kemna, Jan et al.
In: Plant Journal, Vol. 89, No. 1, 14.11.2016, p. 155-168.

Research output: Contribution to journalArticleResearchpeer review

Ordon, J, Gantner, J, Kemna, J, Schwalgun, L, Reschke, M, Streubel, J, Boch, J & Stuttmann, J 2016, 'Generation of chromosomal deletions in dicotyledonous plants employing a user-friendly genome editing toolkit', Plant Journal, vol. 89, no. 1, pp. 155-168. https://doi.org/10.1111/tpj.13319
Ordon, J., Gantner, J., Kemna, J., Schwalgun, L., Reschke, M., Streubel, J., Boch, J., & Stuttmann, J. (2016). Generation of chromosomal deletions in dicotyledonous plants employing a user-friendly genome editing toolkit. Plant Journal, 89(1), 155-168. https://doi.org/10.1111/tpj.13319
Ordon J, Gantner J, Kemna J, Schwalgun L, Reschke M, Streubel J et al. Generation of chromosomal deletions in dicotyledonous plants employing a user-friendly genome editing toolkit. Plant Journal. 2016 Nov 14;89(1):155-168. doi: 10.1111/tpj.13319
Ordon, Jana ; Gantner, Johannes ; Kemna, Jan et al. / Generation of chromosomal deletions in dicotyledonous plants employing a user-friendly genome editing toolkit. In: Plant Journal. 2016 ; Vol. 89, No. 1. pp. 155-168.
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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 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.",
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AU - Ordon, Jana

AU - Gantner, Johannes

AU - Kemna, Jan

AU - Schwalgun, Lennart

AU - Reschke, Maik

AU - Streubel, Jana

AU - Boch, Jens

AU - Stuttmann, Johannes

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