The GE-Qsd1 wheat events were generated by targeted knockout of the homeologous qsd1 loci in Triticum aestivum cv. Fielder using CRISPR/Cas9 via Agrobacterium-mediated transformation.

qsd1 has been shown to regulate seed dormancy levels. The knock-out aims to prevent pre-harvest sprouting of grains on spikes.

A CRISPR/Cas9 construct, with a guide RNAs targeting a conserved region among three sub-genomes in exon 14 of qsd1, was used for transformation of immature wheat embryos.

A triple-knockout mutation of qsd1 was successfully generated.

A transgene-free null-segregant was identified by PCR and whole genome sequencing.

Source: Abe et al. (2019), Cell Rep 28 (5), 1362-1369; doi:10.1016/j.celrep.2019.06.090