Transforming Peanut Plants Using Gene Editing Techniques to Decrease Allergen Content

Peanut allergies pose a significant public health risk, often resulting in severe or life-threatening reactions. This study aimed to reduce the allergenic potential of peanuts by using CRISPR-Cpf1 to target and edit genes responsible for major allergenic proteins. A plasmid named PZJ009 containing the Cpf1 gene and a hygromycin resistance gene cassette were introduced to peanut plants and calluses using both Agrobacterium and biolistic particle delivery approaches. DNA was extracted from the transformed samples and tested with PCR using Cpf1 or hygromycin primers to confirm successful delivery of the plasmid with gel electrophoresis. Out of 54 samples, 2 tested positive for Cpf1 gene, which is a 4% transformation efficiency with the biolistic method being more effective. Further, PCR testing using Ara h primers were done on Cpf1 positive samples to check for presence or absences in the allergen-related gene regions in the plant. While the transformation efficiency was low, this research supports that transformation is possible using gene editing to develop safer, hypoallergenic peanut varieties.