Outlook for coeliac disease patients: towards bread wheat with hypoimmunogenic gluten by gene editing of α- and γ-gliadin gene families

Table 1 Summary of T0 Fielder-CRISPR regenerated plants expressing Cas9 and their nptII copy number

	T0 Fielder wheat plants
	Regenerated	Expressing Cas9	1 or 2 transgene copies
Construct α1_sgRNA sgRNA_α87	40	38 (95%)	21 (55%)
Construct α2_sgRNA sgRNA_α213 + sgRNA_α324	32	12 (38%)	4 (33%)
Construct γ3_sgRNA sgRNA_γ86 + sgRNA_γ272 + sgRNA_γ603	36	35 (97%)	26 (74%)
Construct α2γ3_sgRNA sgRNA_α213 + sgRNA_α324 + sgRNA_γ86 + sgRNA_γ272 + sgRNA_γ603	42	32* (76%)	12 (38%)
TOTAL	150	117 (78%)	63 (54%)

For each construct, between 32 and 42 plants were regenerated. The * indicates that 34 plants expressed Cas9 but 2 were discarded since they did not contain all 5 sgRNAs. Overall, 78% of plants expressed the Cas9, among which 54% had 1 or 2 copies of the nptII selectable marker gene whereas the remainder contained additional copies of nptII.

ISSN: 1471-2229