Document Detail


Alleviating peanut allergy using genetic engineering: the silencing of the immunodominant allergen Ara h 2 leads to its significant reduction and a decrease in peanut allergenicity.
MedLine Citation:
PMID:  17784907     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Peanut allergy is one of the most life-threatening food allergies and one of the serious challenges facing the peanut and food industries. Current proposed solutions focus primarily on ways to alter the immune system of patients allergic to peanut. However, with the advent of genetic engineering novel strategies can be proposed to solve the problem of peanut allergy from the source. The objectives of this study were to eliminate the immunodominant Ara h 2 protein from transgenic peanut using RNA interference (RNAi), and to evaluate the allergenicity of resulting transgenic peanut seeds. A 265-bp-long PCR product was generated from the coding region of Ara h 2 genomic DNA, and cloned as inverted repeats in pHANNIBAL, an RNAi-inducing plant transformation vector. The Ara h 2-specific RNAi transformation cassette was subcloned into a binary pART27 vector to construct plasmid pDK28. Transgenic peanuts were produced by infecting peanut hypocotyl explants with Agrobacterium tumefaciens EHA 105 harbouring the pDK28 construct. A total of 59 kanamycin-resistant peanut plants were regenerated with phenotype and growth rates comparable to wild type. PCR and Southern analyses revealed that 44% of plants stably integrated the transgene. Sandwich ELISA performed using Ara h 2-mAbs revealed a significant (P < 0.05) reduction in Ara h 2 content in several transgenic seeds. Western immunobloting performed with Ara h 2-mAb corroborated the results obtained with ELISA and showed absence of the Ara h 2 protein from crude extracts of several transgenic seeds of the T(0) plants. The allergenicity of transgenic peanut seeds expressed as IgE binding capacity was evaluated by ELISA using sera of patients allergic to peanut. The data showed a significant decrease in the IgE binding capacity of selected transgenic seeds compared to wild type, hence, demonstrating the feasibility of alleviating peanut allergy using the RNAi technology.
Authors:
Hortense W Dodo; Koffi N Konan; Fur C Chen; Marceline Egnin; Olga M Viquez
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Publication Detail:
Type:  Journal Article     Date:  2007-09-03
Journal Detail:
Title:  Plant biotechnology journal     Volume:  6     ISSN:  1467-7652     ISO Abbreviation:  Plant Biotechnol. J.     Publication Date:  2008 Feb 
Date Detail:
Created Date:  2008-01-11     Completed Date:  2008-08-07     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  101201889     Medline TA:  Plant Biotechnol J     Country:  England    
Other Details:
Languages:  eng     Pagination:  135-45     Citation Subset:  IM    
Affiliation:
Department of Food and Animal Sciences, Food Biotechnology Laboratory, Alabama A&M University, Normal, AL 35762, USA. hortense.dodo@email.aamu.eduhortense.dodo@email.aamu.edu
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MeSH Terms
Descriptor/Qualifier:
2S Albumins, Plant
Allergens / genetics,  immunology*
Blotting, Southern
Enzyme-Linked Immunosorbent Assay
Gene Silencing*
Genetic Engineering*
Glycoproteins / genetics,  immunology*
Humans
Immunoglobulin E / blood
Kanamycin Resistance
Peanut Hypersensitivity / prevention & control,  therapy*
Plant Proteins / genetics,  immunology*
Plants, Genetically Modified
Polymerase Chain Reaction
RNA Interference
Chemical
Reg. No./Substance:
0/2S Albumins, Plant; 0/Allergens; 0/Ara h 2 allergen, Arachis hypogaea; 0/Glycoproteins; 0/Plant Proteins; 37341-29-0/Immunoglobulin E

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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