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. 2021 Jun 22;12:3854. doi: 10.1038/s41467-021-24195-5

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a A schematic representation of a non-autonomous trans-acting gene drive. A Cas9 unit and a gRNA unit are placed in a location far away from the target gene (on a different chromosome). The Cas9/gRNA complex cuts the wild type allele of the target gene. Subsequently, the DSB is repaired using the DNA from the homologous chromosome as the template, resulting in a homozygous plant. b The NPY5-GFP line can be used to test a non-autonomous trans-acting gene drive in Arabidopsis. The line contains an EC-Cas9 unit and a gRNA unit on Chromosome II while the target wild type NPY5 is on Chromosome IV. The NPY5 gene has been tagged with GFP. The DD45-Cas9 unit on Chromosome V may provide additional Cas9 for the non-autonomous trans-acting gene drive. c The functionality of the non-autonomous trans-acting gene drive was tested by crossing the NPY5-GFP line to wild type Ler. Cas9/gRNA from Chromosome II cuts wild type NPY5, generating a DSB. The GFP and DNA flanking GFP is used as a template for HDR, resulting in homozygous NPY5-GFP F1 plants.