GE-FAD2KO Soybean line GM026-18 is a transgene-free knock-out line with mutations in both the FAD2-1A and the FAD2-1B gene.
The fatty acid desaturase 2 (FAD2) determines the level of monounsaturated fats in soybean oil and vegetative tissue. The FAD2KO Soybean knock-out line is homozygous for mutations in both FAD2 genes resulting elevated levels of oleic acid that approximate 80 %.
GE-FAD2KO Soybean line GM026-18 was developed by Agrobacterium rhizogenes-mediated hairy-root transformation of soybean cotyledones of the variety Bert with a TALEN T-DNA expression construct to knock-out FAD2.
The sequence-specific TALENs target all alleles of the FAD2-1A and FAD2-1B genes of soybean. In both genes the target sites are located within 500 bp of the start codon.
The expression constructs consisted of the 35S promoter, two TALEN monomer coding sequences linked by a T2A sequence, and a nos terminator.
PCR analyses targeting three distinct regions of the T-DNA construct were used to determine that the TALEN T-DNA construct was segregated away.
Both Haun et al. (2014) Plant Biotech J. 2:934-940 and the Patent WO2014141147A1 describe the line GM026-18. The 2014 “Am I regulated” Inquiry of the company Cellectis for FAD2KO Soybean is less specific with regard to the specific line.