title: Dau c 1.01 and Dau c 1.02-silenced transgenic carrot plants show reduced allergenicity to patients with carrot allergy.
abstract: Pathogenesis-related protein-10 (PR10) is a ubiquitous small plant protein induced by microbial pathogens and abiotic stress that adversely contributes to the allergenic potency of many fruits and vegetables, including carrot. In this plant, two highly similar genes encoding PR10 isoforms have been isolated and designated as allergen Dau c 1.01 and Dau c 1.02. The aim of the study was to generate PR10-reduced hypoallergenic carrots by silencing either one of these genes in transgenic carrots by means of RNA interference (RNAi). The efficiency of gene silencing by stably expressed hairpin RNA (hnRNA) was documented by means of quantitative RT-PCR (qPCR) and immunoblotting. Quantification of the residual protein revealed that PR10 accumulation was strongly decreased compared with untransformed controls. Treatment of carrot plants with the PR protein-inducing chemical salicylic acid resulted in an increase of PR10 isoforms only in wild-type but not in Dau c 1-silenced mutants. The decrease of the allergenic potential in Dau c 1-silenced plants was sufficient to cause a reduced allergenic reactivity in patients with carrot allergy, as determined with skin prick tests (SPT). However, simultaneous silencing of multiple allergens will be required to design hypoallergenic carrots for the market. Our findings demonstrate the feasibility of creating low-allergenic food by using RNAi. This constitutes a reasonable approach to allergen avoidance.
authors.affiliation: Research Centre for BioSystems, Land Use and Nutrition, Justus Liebig University, Giessen, Germany.
authors.forename: Susanna
authors.initials: S
authors.lastname: Peters
authors.forename: Jafargholi
authors.initials: J
authors.lastname: Imani
authors.forename: Vera
authors.initials: V
authors.lastname: Mahler
authors.forename: Kay
authors.initials: K
authors.lastname: Foetisch
authors.forename: Susanne
authors.initials: S
authors.lastname: Kaul
authors.forename: Kathrin E
authors.initials: KE
authors.lastname: Paulus
authors.forename: Stephan
authors.initials: S
authors.lastname: Scheurer
authors.forename: Stefan
authors.initials: S
authors.lastname: Vieths
authors.forename: Karl-Heinz
authors.initials: KH
authors.lastname: Kogel
mesh_terms: D052179:Antigens, Plant / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D018552:Daucus carota / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D005512:Food Hypersensitivity / Q000276:immunology*; D030841:Food, Genetically Modified; D020868:Gene Silencing; D006801:Humans; D015151:Immunoblotting; D007073:Immunoglobulin E / Q000097:blood; D009154:Mutation; D010940:Plant Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D020033:Protein Isoforms / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012882:Skin Tests
chemical_list: D052179:Antigens, Plant; C119925:Dau c 1 protein, Daucus carota; D010940:Plant Proteins; D020033:Protein Isoforms; D034741:RNA, Small Interfering; D007073:Immunoglobulin E
references.citation: Plant J. 2008 Feb;53(4):674-90
references.pmid: 18269576
references.citation: Ann N Y Acad Sci. 2002 May;964:47-68
references.pmid: 12023194
references.citation: Eur J Biochem. 2001 Oct;268(20):5310-20
references.pmid: 11606193
references.citation: Cell Microbiol. 2005 Nov;7(11):1555-64
references.pmid: 16207243
references.citation: Clin Exp Allergy. 2005 Oct;35(10):1392-9
references.pmid: 16238801
references.citation: Plant Physiol. 2003 May;132(1):36-43
references.pmid: 12746509
references.citation: Mol Plant Pathol. 2010 Jan;11(1):69-81
references.pmid: 20078777
references.citation: J Allergy Clin Immunol. 2001 Aug;108(2):301-7
references.pmid: 11496252
references.citation: Clin Exp Allergy. 2005 Jul;35(7):970-8
references.pmid: 16008686
references.citation: Mol Plant Pathol. 2002 May 1;3(3):135-44
references.pmid: 20569319
references.citation: Annu Rev Phytopathol. 2006;44:135-62
references.pmid: 16602946
references.citation: FEBS Lett. 1996 Aug 12;391(3):341-5
references.pmid: 8765003
references.citation: J Allergy Clin Immunol. 2005 Feb;115(2):364-9
references.pmid: 15696096
references.citation: Plant Biotechnol J. 2006 Mar;4(2):231-42
references.pmid: 17177799
references.citation: Clin Exp Allergy. 1999 Jun;29(6):840-7
references.pmid: 10336602
references.citation: J Allergy Clin Immunol. 2005 Jul;116(1):213-9
references.pmid: 15990797
references.citation: Exp Cell Res. 1968 Apr;50(1):151-8
references.pmid: 5650857
references.citation: Anal Biochem. 1976 May 7;72:248-54
references.pmid: 942051
references.citation: Trends Plant Sci. 2002 May;7(5):193-5
references.pmid: 11992820
references.citation: Biotechnology (N Y). 1991 Oct;9(10):963-7
references.pmid: 1368724
references.citation: Clin Exp Allergy. 2002 Sep;32(9):1329-40
references.pmid: 12220472
references.citation: J Plant Physiol. 2010 Jan 15;167(2):121-30
references.pmid: 19682768
references.citation: Planta. 2007 Jun;226(1):57-71
references.pmid: 17237953
references.citation: Biosci Biotechnol Biochem. 1996 Aug;60(8):1215-21
references.pmid: 8987539