CRISPR-Cas9 Used for Targeted Disruption of FAD2–2 Microsomal Omega-6 Desaturase in Soybean

RNA-guided CRISPR-Cas9 system has been regarded as a useful tool in site-directed mutagenesis because of its wide applications in various branches of biology. CRISPR-Cas9 nuclease system displaced the previously used tools and their flaws in site-specific mutagenesis. Thus, this system was used by Piwu Wang of Jilin Agricultural University and team and showed an efficient sequence specific integration/mutation of FAD2–2 gene in soybean. The results of their study was published in BMC Biotechnology.

The researchers designed a single guided RNA sequence using bioinformatics tools aimed at targeting specific portions of the FAD2-2 loci in soybean, which is vital in the biosynthesis of peak oil. Findings showed that the soybean transgenic mutants subjected to targeted mutation were detected in the target gene. Furthermore, the detection of Cas9 gene, BAR gene, and NOS terminator were carried out respectively. Further analysess confirmed the stable transformation of Cas9 gene into soybean, as well as the efficient CRISPR-Cas9/sgRNA induced mutation within the target sequence of FAD2–2 loci. FAD2–2 integration at target region in the form of substitution, deletions, and insertions were completed with notably high frequency and rare off-target mutagenesis.

The findings provides a summary of recent developments and future challenges in producing agronomically important crops using the CRISPR-Cas9 system.

Read the research article from BMC Biotechnology.