GE-BADC Camelina lines were developed by CRISPR/Cas9 technology targeting the endogenous badc genes.
The Biotin Attachment Domain-Containing (BADC) protein is a negative regulator of the enzyme acetyl-CoA carboxylase (ACCase), the key enzyme for fatty acid biosynthesis. Its normal function slows down oil biosynthesis.
The intended phenotype of GE-BADC Camelina lines is an increased amount of oil in Camelina seed via inactivation of the three homeologs of the badc gene.
A binary vector with expression cassettes encoding the endonuclease Cas9 and three guide RNAs to target the endogenous badc genes was delivered into the plant cells by Agrobacterium-mediated transformation and the floral dip method.
DNA sequence analysis has shown that the GE-BADC Camelina lines contain two to twenty-two nucleotide deletions that either disrupt the coding sequence of the badc genes, or that delete one codon from the genes.
The absence of transgenic insertions in null segregant GE-BADC Camelina lines was confirmed by PCR analysis.