@article{YanChenKaufmann2016,
  author    = {Yan, Wenhao and Chen, Dijun and Kaufmann, Kerstin},
  title     = {Efficient multiplex mutagenesis by RNA-guided Cas9 and its use in the characterization of regulatory elements in the AGAMOUS gene},
  series = {Plant Methods},
  volume    = {12},
  journal   = {Plant Methods},
  publisher = {BioMed Central},
  address   = {London},
  issn      = {1746-4811},
  doi       = {10.1186/s13007-016-0125-7},
  pages     = {2381 -- 2389},
  year      = {2016},
  abstract  = {Results: Here, an RNA-guided Cas9 system was optimized to enable efficient multiplex editing in Arabidopsis thaliana. We demonstrate the flexibility of our system for knockout of multiple genes, and to generate heritable large-fragment deletions in the genome. As a proof of concept, the function of part of the second intron of the flower development gene AGAMOUS in Arabidopsis was studied by generating a Cas9-free mutant plant line in which part of this intron was removed from the genome. Further analysis revealed that deletion of this intron fragment results 40 \% decrease of AGAMOUS gene expression without changing the splicing of the gene which indicates that this regulatory region functions as an activator of AGAMOUS gene expression. Conclusions: Our modified RNA-guided Cas9 system offers a versatile tool for the functional dissection of coding and non-coding DNA sequences in plants.},
  language  = {en}
}
@article{YanChenKaufmann2016,
  author    = {Yan, Wenhao and Chen, Dijun and Kaufmann, Kerstin},
  title     = {Efficient multiplex mutagenesis by RNA-guided Cas9 and its use in the characterization of regulatory elements in the AGAMOUS gene},
  series = {Plant methods},
  volume    = {12},
  journal   = {Plant methods},
  publisher = {BioMed Central},
  address   = {London},
  issn      = {1746-4811},
  doi       = {10.1186/s13007-016-0125-7},
  pages     = {1 -- 9},
  year      = {2016},
  abstract  = {Background The efficiency of multiplex editing in plants by the RNA-guided Cas9 system is limited by efficient introduction of its components into the genome and by their activity. The possibility of introducing large fragment deletions by RNA-guided Cas9 tool provides the potential to study the function of any DNA region of interest in its 'endogenous' environment. Results Here, an RNA-guided Cas9 system was optimized to enable efficient multiplex editing in Arabidopsis thaliana. We demonstrate the flexibility of our system for knockout of multiple genes, and to generate heritable large-fragment deletions in the genome. As a proof of concept, the function of part of the second intron of the flower development gene AGAMOUS in Arabidopsis was studied by generating a Cas9-free mutant plant line in which part of this intron was removed from the genome. Further analysis revealed that deletion of this intron fragment results 40 \% decrease of AGAMOUS gene expression without changing the splicing of the gene which indicates that this regulatory region functions as an activator of AGAMOUS gene expression. Conclusions Our modified RNA-guided Cas9 system offers a versatile tool for the functional dissection of coding and non-coding DNA sequences in plants.},
  language  = {en}
}