SBIR Phase I: Progeny Analysis Of A Chimera-Free Hypoallergenic Peanut

这项小企业创新研究（SBIR）一期项目的更广泛影响/商业潜力是开发一种低过敏性花生（HAP）品种，该品种有望通过预防与过敏相关的死亡来挽救人类生命，减少花生过敏的发生率，并为那些对花生敏感的个人及其家人、大家庭、朋友、学校和餐馆增加安全、舒适和安心。HAP还可以减少和/或消除由花生过敏事件引起的机构诉讼的数量，减少和/或消除因花生污染而导致的昂贵的食品召回的数量。减少过敏原可能会挽回花生市场遭受的损失，提高美国在国际花生市场上的竞争优势，并由于餐馆、航空公司等市场的增加而使销售额增加15%至20%。食物过敏每年造成250亿美元的损失，花生是其中的主要原因。HAP将为花生过敏提供一个重要的解决方案，有可能降低食物过敏的总体成本。这个SBIR一期项目的智力价值在于生产一种低过敏性花生的商业原型，以解决全国范围内花生过敏的严重健康问题。在先前的工作中，通过同时沉默三种最有效的花生过敏原Ara h1、Ara h2和Ara h3的编码基因，开发了一种HAP。在田间试验中，HAP的致敏效力降低了2 ~ 85倍，并连续几代遗传。然而，一些转基因花生被发现是嵌合的，产生的种子群体不均匀，具有不同程度的致敏效力。为了解决这一问题，重复体细胞胚性组织被用于产生第一代无嵌合体的HAP （T0植株）。在第一期项目中，将使用Southern blot和定量PCR对后代T1植株进行分析，以确定植株的无嵌合体状态。将进行逆转录酶定量PCR、Western blot、夹心ELISA和免疫帽检测，以确认后代T2种子的群体是同质的，并且目标过敏原的水平是检测不到的。低过敏性花生的商业原型品种将被确定并选择用于进一步的研究。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值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/commercial potential of this Small Business Innovation Research (SBIR) Phase I project is to develop a hypo allergenic peanut (HAP) variety, which is expected to save human lives by preventing allergy-related deaths, reduce the incidence of peanut allergy, and increase security, comfort and peace of mind for those individuals sensitive to peanuts and their families, extended families, friends, schools, and restaurants. HAP also may reduce and/or eliminate the number of institutional lawsuits resulting from peanut allergy events, and reduce and/or eliminate the number of costly food recalls due to peanut contaminations. Reducing allergens may gain back the loss endured by the peanut market, improve the U.S. competitive edge on the international peanut market, and increase sales by 15% to 20% due to an increase market by restaurants, airlines and more. Food allergies cost the US $25 billion annually and peanut is a major contributor. HAP will provide a significant solution to peanut allergy with the potential to reduce the overall cost of food allergy. The intellectual merit of this SBIR Phase I project is to produce a commercial prototype for a hypoallergenic peanut as a solution to the serious health concern nationwide for peanut allergy. In previous work, a HAP was developed by concomitant silencing of the genes coding for the three most potent peanut allergens Ara h 1, Ara h 2 and Ara h 3. The HAP was 2 to 85-fold reduced in allergenic potency, and the HAP trait was transmitted to several successive generations in field trials. However, some of the transgenic peanuts were found to be chimeric and to produce non-homogeneous population of seeds with variable levels of allergenic potency. To solve this problem, repetitive somatic embryogenic tissue was used to produce first generation chimera-free HAP (T0 plants). During this Phase I project, the progeny T1 plants will be analyzed using Southern blot and quantitative PCR to confirm the chimera-free status of the plants. Reverse transcriptase quantitative PCR, Western blot, sandwich ELISA and immunoCap assay will be performed to confirm the population of the progeny T2 seeds is homogenous and has undetectable levels of the targeted allergens. A commercial prototype variety for the hypoallergenic peanut will be identified and selected for further studies.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.