One-step genome editing of elite crop germplasm during haploid induction.

Kelliher, Timothy; Starr, Dakota; Su, Xiujuan; Tang, Guozhu; Chen, Zhongying; Carter, Jared; Wittich, Peter E; Dong, Shujie et al. · Nat Biotechnol · 2019

basic_science · Level V

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Abstract

Genome editing using CRISPR-Cas9 works efficiently in plant cells<sup>1</sup>, but delivery of genome-editing machinery into the vast majority of crop varieties is not possible using established methods<sup>2</sup>. We co-opted the aberrant reproductive process of haploid induction (HI)<sup>3-6</sup> to induce edits in nascent seeds of diverse monocot and dicot species. Our method, named HI-Edit, enables direct genomic modification of commercial crop varieties. HI-Edit was tested in field and sweet corn using a native haploid-inducer line<sup>4</sup> and extended to dicots using an engineered CENH3 HI system<sup>7</sup>. We also recovered edited wheat embryos using Cas9 delivered by maize pollen. Our data indicate that a transient hybrid state precedes uniparental chromosome elimination in maize HI. Edited haploid plants lack both the haploid-inducer parental DNA and the editing machinery. Therefore, edited plants could be used in trait testing and directly integrated into commercial variety development.

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