A genetic engineering strategy to eliminate peanut allergy

Abstract

Peanut allergy is an IgE-mediated hypersensitivity reaction with an increasing prevalence worldwide. Despite its seriousness, to date, there is no cure. Genetic engineering strategies can provide a solution. The post-transcriptional gene silencing (PTGS) model can be used effectively to knock out the production of allergenic proteins in peanut by specific degradation of the endogenous target messenger RNA (mRNA). Ara h 2, the most potent peanut allergenic protein, was selected as a model to demonstrate the feasibility of this concept. Transgenic peanut plants were produced via microprojectile-mediated transformation of peanut embryos using a plasmid construct, which contains a fragment of the coding region of Ara h 2 linked to an enhanced CaMV 35S constitutive promoter. Molecular analyses, including polymerase chain reaction and Southern blots, confirmed the presence of the stable integration of the Ara h 2 transgene into the peanut genome. Northern hybridization showed the expression of the Ara h 2 transgene in all vegetative tissues of the mature transgenic peanut plants, indicating the stable expression of the truncated Ara h 2 transgene throughout the development of the plants. It is, therefore, reasonable to expect that the truncated Ara h 2 transgene transcripts will be synthesized in the seeds and will trigger the specific degradation of endogenous Ara h 2 mRNA. The next step will be to grow the transgenic peanut plants to full maturity for seed production and to determine the level of allergen Ara h 2.

Skin prick tests reveal stable and heritable reduction of allergenic potency of gene-silenced tomato fruits

“…Recently, the RNAi technology was applied to suppress the expression of the major apple allergen Mal d 1 3 and Ara h 2 in peanuts. 4 Preliminary data showed suppression of Ara h 2 in plantlets, as identified by means of PCR and Southern and Northern blotting. Moreover, suppression of Mal d 1 was analyzed by using IgE and IgG immunoblotting.…”

Detection of Antigens Reactive to IgE and IgA during Wheat Seed Maturation and in Different Wheat Cultivars

“…Natural breeding and breeding in combination with random mutation strategies [38,39,40] accompanied by serological analysis may allow us to foster effectively viable hypoallergenic cultivars. Several attempts have been made so far to silence the expression of certain allergens in rice, grass pollen, apple and tomato by anti-sense technologies [23,24,25,26,27,28,29]. Using this technology it has been possible to down-regulate the expression of certain allergens but it may affect growth and viability of the engineered plants.…”

“…Preventative research has included silencing allergenic proteins in plants by anti-sense technology, which has been used to reduce the expression of certain allergens in rice, grass pollen, apples and tomatoes, with the aim of reducing their allergenic activity [23,24,25,26,27,28,29]. …”

Reduction of Allergenic Proteins by the Effect of theripening inhibitor(rin) Mutant Gene in an F1Hybrid of therinMutant Tomato

“…[31][32][33][34][35] Although these approaches are highly effective in eliminating a target protein, they are limited to only one allergen. The result of this study, in contrast, suggests that multiple allergenic proteins accumulated at a significantly low level in the rin hybrid fruit (Fig.…”