4.8 Article

Removing the major allergen Bra j I from brown mustard (Brassica juncea) by CRISPR/Cas9

Journal

PLANT JOURNAL
Volume 109, Issue 3, Pages 649-663

Publisher

WILEY
DOI: 10.1111/tpj.15584

Keywords

mustard; Bra j I; CRISPR; Cas; transformation; food allergen; seed storage protein

Categories

Funding

  1. Federal Ministry of Education and Research (BMBF LACOP consortium) [031B0348]
  2. Projekt DEAL

Ask authors/readers for more resources

The study successfully demonstrated the potential of gene editing breeding to alleviate food allergies by removing or mutating the major allergen Bra j I in brown mustard using CRISPR/Cas9 technology.
Food allergies are a major health issue worldwide. Modern breeding techniques such as genome editing via CRISPR/Cas9 have the potential to mitigate this by targeting allergens in plants. This study addressed the major allergen Bra j I, a seed storage protein of the 2S albumin class, in the allotetraploid brown mustard (Brassica juncea). Cotyledon explants of an Indian gene bank accession (CR2664) and the German variety Terratop were transformed using Agrobacterium tumefaciens harboring binary vectors with multiple single guide RNAs to induce either large deletions or frameshift mutations in both Bra j I homoeologs. A total of 49 T-0 lines were obtained with up to 3.8% transformation efficiency. Four lines had large deletions of 566 up to 790 bp in the Bra j IB allele. Among 18 Terratop T-0 lines, nine carried indels in the targeted regions. From 16 analyzed CR2664 T-0 lines, 14 held indels and three had all four Bra j I alleles mutated. The majority of the CRISPR/Cas9-induced mutations were heritable to T-1 progenies. In some edited lines, seed formation and viability were reduced and seeds showed a precocious development of the embryo leading to a rupture of the testa already in the siliques. Immunoblotting using newly developed Bra j I-specific antibodies revealed the amount of Bra j I protein to be reduced or absent in seed extracts of selected lines. Removing an allergenic determinant from mustard is an important first step towards the development of safer food crops.

Authors

I am an author on this paper
Click your name to claim this paper and add it to your profile.

Reviews

Primary Rating

4.8
Not enough ratings

Secondary Ratings

Novelty
-
Significance
-
Scientific rigor
-
Rate this paper

Recommended

No Data Available
No Data Available