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General Information

Document type
  • Peer-reviewed journal article
GE organism
  • apple
GE trait
  • quality
Country
  • The Netherlands, Austria

Results

Efficacy
  • positive effect
Safety for consumption
  • positive effect

Silencing the major apple allergen Mal d 1 by using the RNA interference approach Open Access

Gilissen, LJ; Bolhaar, ST; Matos, CI; Rouwendal, GJ; Boone, MJ; Krens, FA; Zuidmeer, L; Van Leeuwen, A; Akkerdaas, J; Hoffmann-Sommergruber, K; Knulst, AC; Bosch, D; Van de Weg, WE; Van Ree, R
The Journal of Allergy and Clinical Immunology. 2005 February. 115(2):364-369

Link to full text (open access, freely available)

PMID: 15696096 DOI: 10.1016/j.jaci.2004.10.014 ISSN: 0091-6749

Abstract

BACKGROUND: Apple allergy is dominated by IgE antibodies against Mal d 1 in areas where birch pollen is endemic. Apples with significantly decreased levels of Mal d 1 would allow most patients in these areas to eat apples without allergic reactions. OBJECTIVE: The aim of this study was to inhibit the expression of Mal d 1 in apple plants by RNA interference. METHODS: In vitro -grown apple plantlets were transformed with a construct coding for an intron-spliced hairpin RNA containing a Mal d 1-specific inverted repeat sequence separated by a Mal d 1-specific intron sequence. The presence of the construct in transformants was checked by PCR. Expression of Mal d 1 in leaves was monitored by prick-to-prick skin testing in 3 patients allergic to apples and by immunoblotting with a Mal d 1-reactive mAb and with IgE antibodies against Mal d 1. RESULTS: After transformation, plantlets were selected on the basis of having a normal phenotype and growth rate. With PCR, in 6 of 9 selected plantlets, the presence of the gene-silencing construct was demonstrated. By skin prick test it was shown that a wild-type plantlet had significantly ( P < .05) higher allergenicity than 5 of the transformants. Reduction of expression of Mal d 1 was confirmed by immunoblotting. In wild-type and unsuccessful transformants, a strong band was detected with Mal d 1-reactive mAb 5H8 at the expected apparent M r of 17 kDa. This band was virtually absent in the transformants that carried the gene-silencing construct. With human IgE antibodies, the same observations were made. CONCLUSIONS: Mal d 1 expression was successfully reduced by RNA interference. This translated into significantly reduced in vivo allergenicity. These observations support the feasibility of the production by gene silencing of apples hypoallergenic for Mal d 1.

Keywords

Gene silencing, Mal d 1, RNA interference, genetic modification, apple (Malus domestica), skin prick test, hypoallergenic, immunoblotting

Funding

Funding source
  • EU-SAFE Project
  • Duch Ministry of Agriculture, Nature Management and Food Safety
Funding country
  • European Union
  • The Netherlands
Funding type
  • government

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Cite this study

MLA

Gilissen, LJ, ST Bolhaar, CI Matos, GJ Rouwendal, MJ Boone, FA Krens, L Zuidmeer, A Van Leeuwen, J Akkerdaas, K Hoffmann-Sommergruber, AC Knulst, D Bosch, WE Van de Weg, R Van Ree. "Silencing the major apple allergen Mal d 1 by using the RNA interference approach." The Journal of Allergy and Clinical Immunology 115.2 (2005): 364-369. Web. 20 Nov. 2017.

APA

Gilissen, LJ., Bolhaar, ST., Matos, CI., Rouwendal, GJ., Boone, MJ., Krens, FA., Zuidmeer, L., Van Leeuwen, A., Akkerdaas, J., Hoffmann-Sommergruber, K., Knulst, AC., Bosch, D., Van de Weg, WE., Van Ree, R. (2005). Silencing the major apple allergen Mal d 1 by using the RNA interference approach. The Journal of Allergy and Clinical Immunology, 115(2), 364-369. doi:10.1016/j.jaci.2004.10.014

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