The GE-DGAT1B soybean line was generated by CRISPR-Cas editing of the diacylglycerol acyltransferase (dgat1) gene in Glycine max.

The dgat1b gene encodes the type 1 diacylglycerol acyltransferases which catalyzes the final step in oil (triacylglycerol) production.

Aim of the mutation of the dgat1b gene was to improve the kinetic properties of the DGAT1 enzyme in order to increase oil and protein content in the soybean line.

CRISPR-Cas9 gene editing was used to introduce the targeted base pair substitutions of the dgat1b gene. The components of the gene editing system were delivered via Ochrobactrum haywardense strain H1-8 mediated transformation with one plasmid.

The intended base pair substitutions were confirmed by Next Generation Sequencing (NGS) analysis. The GE-DGAT1B soybean line contains a single amino acid substitution, Isoleucine (I) to Serine (S), at position 479 (I479S).

Southern-by-sequencing was used to confirm absence of unintentionally integrated DNA from the transformation plasmid in the final soybean line.

Field trials are intended to evaluate the efficacy of the introduced mutation.

Source:

USDA APHIS letter of inquiry