SBIR Phase II: Development of An Allergen-Free Peanut Using Genome Editing Technology

这个小企业创新研究(SBIR)第二阶段项目的更广泛影响是解决与花生过敏相关的健康问题。花生过敏影响了大量的人群，对那些病情最严重的人来说可能是极其危险的。鉴于花生产品的无处不在，花生过敏是世界范围内一个重大的医学和法律问题，这种潜在的致命疾病在儿童中的发病率不断上升。由该项目开发的无过敏原花生具有显著降低花生威胁生命的反应的潜力，以及以下好处：(1)降低食物过敏的总体发生率，(2)通过消除因意外摄入花生而导致的过敏反应和死亡来保护人类的生命，(3)消除花生过敏者及其家人的情绪困扰，(4)增强公众对花生作为健康促进食品的认知，以及(5)减少因花生污染而导致的产品召回次数。拟议的项目旨在利用CRISPR/CAS(成簇的规则间隔短回文重复序列和CRISPR相关系统)等基因组编辑工具开发一种不含所有临床记录的过敏原的花生并将其商业化。花生过敏原与其已与肥大细胞结合的特异性IgE抗体接触后不久就会发生花生过敏反应。花生变应原特异性IgE的交联会刺激肥大细胞释放与临床症状有关的化学介质，有时包括过敏性休克或死亡。阿拉伯h1、阿拉伯h2、阿拉伯h3和阿拉伯h6是临床上记录最多的花生过敏原。该项目的目标是：(1)构建和验证ara h基因编辑载体，转化花生胚性愈伤组织并再生T0花生植株；(2)鉴定ara h基因缺失的T0个个体；(3)组装携带所有目标ara h基因纯合子缺失的品系；(5)鉴定所选品系的ara h基因和蛋白表达；(6)评估温室条件下无Arah品系的营养发育。如果由此产生的植物表现出对基因缺失的耐受性，无过敏原花生将在保护花生敏感人群和促进花生销售方面具有非常重要的意义。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact of this Small Business Innovation Research (SBIR) Phase II project is to address health concerns associated with peanut allergies. Peanut allergies affect a large population and can be extremely dangerous for those with the most severe forms. Given the ubiquity of peanut products, peanut allergy is a significant medical and legal concern worldwide, with a rising incidence of this potentially fatal condition in children. An allergen-free peanut developed from this project has the potential to significantly dampen the life-threatening reactions to peanuts, as well as the following benefits: (1) reducing the overall incidence of food allergy, (2) preserving human lives by eliminating anaphylaxis and death caused by accidental ingestion of peanut, (3) eliminating emotional distress of peanut allergic individuals and their families, (4) enhancing the public perception of peanuts as a health-promoting food, and (5) reducing the number of product recalls due to peanut contamination. The proposed project aims to develop and to commercialize an allergen-free peanut devoid of all clinically documented allergens using a genome-editing tool such as CRISPR/Cas (clustered regularly interspaced short palindromic repeats and CRISPR-associated system). Allergic reactions to peanut occur shortly after contact of peanut allergens with their specific IgE antibodies already bound to mast cells. The crosslinking of allergen specific IgE by the respective peanut allergens stimulates mast cells to release chemical mediators responsible for the clinical symptoms including sometime anaphylactic shock or death. Ara h 1, Ara h 2, Ara h 3 and Ara h 6 are the most clinically documented peanut allergens. The objectives of the project are: (1) Construct and validate vectors for Ara h gene editing, transform peanut embryogenic callus and regenerate T0 peanut plants; (2) Identify T0 individuals with deletions in Ara h genes; (3) Assemble lines carrying homozygous deletions in all target Ara h genes; (5) Characterize the selected lines for Ara h mRNA and protein expression; (6) Assess vegetative development of Arah-free lines under greenhouse conditions. If the resulting plants demonstrate tolerance to the gene deletions, the allergen-free peanut will be very significant in protecting the peanut sensitive populations and promoting peanut sales.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.