CRISPR/Cas9 Gene Editing of Gluten in Wheat to Reduce Gluten Content and Exposure—Reviewing Methods to Screen for Coeliac Safety

Jouanin, Aurélie; Gilissen, L.J.W.J.; Schaart, Jan G.; Leigh, Fiona; Cockram, James; Wallington, Emma J.; Boyd, Lesley A.; Broeck, H.C. van den; Meer, I.M. van der; America, A.H.P.; Visser, Richard G. F.; Smulders, M.J.M.

doi:10.3389/fnut.2020.00051

Cited by 67 publications

(37 citation statements)

References 104 publications

(152 reference statements)

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“…This T‐cell epitope has been shown to make only a minor contribution to the gluten‐induced T‐cell response in HLA‐DQ2+ CD (Anderson et al ., 2006). These accessions can be used to generate less toxic synthetic hexaploid wheat (SHW) lines, by hybridising these accessions with tetraploid wheat accessions, which would form a good basis for of a breeding programme to generate hypoimmunogenic or even CD‐safe bread wheat (Jouanin et al ., 2018, 2020b). The tetraploid wheat accessions may be selected based on genetic characteristics or agronomic performance.…”

Section: Discussionmentioning

confidence: 99%

Exploring the alpha‐gliadin locus: the 33‐mer peptide with six overlapping coeliac disease epitopes in Triticum aestivum is derived from a subgroup of Aegilops tauschii

et al. 2021

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Summary Most alpha‐gliadin genes of the Gli‐D2 locus on the D genome of hexaploid bread wheat (Triticum aestivum) encode for proteins with epitopes that can trigger coeliac disease (CD), and several contain a 33‐mer peptide with six partly overlapping copies of three epitopes, which is regarded as a remarkably potent T‐cell stimulator. To increase genetic diversity in the D genome, synthetic hexaploid wheat lines are being made by hybridising accessions of Triticum turgidum (AB genome) and Aegilops tauschii (the progenitor of the D genome). The diversity of alpha‐gliadins in A. tauschii has not been studied extensively. We analysed the alpha‐gliadin transcriptome of 51 A. tauschii accessions representative of the diversity in A. tauschii. We extracted RNA from developing seeds and performed 454 amplicon sequencing of the first part of the alpha‐gliadin genes. The expression profile of allelic variants of the alpha‐gliadins was different between accessions, and also between accessions of the Western and Eastern clades of A. tauschii. Generally, both clades expressed many allelic variants not found in bread wheat. In contrast to earlier studies, we detected the 33‐mer peptide in some A. tauschii accessions, indicating that it was introduced along with the D genome into bread wheat. In these accessions, transcripts with the 33‐mer peptide were present at lower frequencies than in bread wheat varieties. In most A. tauschii accessions, however, the alpha‐gliadins do not contain the epitope, and this may be exploited, through synthetic hexaploid wheats, to breed bread wheat varieties with fewer or no coeliac disease epitopes.

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Section: Discussionmentioning

confidence: 99%

Exploring the alpha‐gliadin locus: the 33‐mer peptide with six overlapping coeliac disease epitopes in Triticum aestivum is derived from a subgroup of Aegilops tauschii

et al. 2021

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“… 131 RNA interference (RNAi) and CRISPR/Cas9 are the two recent biotechnology methods used in this regard. 126 …”

Section: Methodsmentioning

confidence: 99%

“…15,27,125 As a result, several genetic studies have focused on breeding wheat with low immunological properties and preserved baking quality through biotechnological approaches. Numerous scientists believe that gene editing would be a definitive solution for GRDs; however, due to the variety of causal agents, it is not so easy to solve them all in this way [126][127][128] Biotechnological approaches are used for precise and organized modifying of specific genomic sequences through their different functions such as gene replacement, targeted gene knock-out and knockin, etc. 129, 130 Vasil et al 131 were the first group to successfully produce transgenic wheat plants in 1992 through the Bar gene transferring by biolistic particle bombardment method.…”

Section: Wheat Genome Editingmentioning

confidence: 99%

“…131 RNA interference (RNAi) and CRISPR/Cas9 are the two recent biotechnology methods used in this regard. 126 RNA interference (RNAi) is a post-transcriptional process present in almost all eukaryotic organisms and regulates the expression of protein-coding genes in a sequence-specific manner, which is capable of engineering novel phenotypes. 132,133 In fact, RNAi suppresses protein synthesis by using short double-stranded RNA (dsRNA) complementary to target mRNA and degrading that (silencing of the gene).…”

Section: Wheat Genome Editingmentioning

confidence: 99%

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The Gluten Gene: Unlocking the Understanding of Gluten Sensitivity and Intolerance

Asri¹,

Rostami-Nejad²,

Rostami³

2021

TACG

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Wheat flour is one of the most important food ingredients containing several essential nutrients including proteins. Gluten is one of the major protein components of wheat consisted of glutenin (encoded on chromosome 1) and gliadin (encoded on chromosome 1 and 6) and there are around hundred genes encoding it in wheat. Gluten proteins have the ability of eliciting the pathogenic immune responses and hypersensitivity reactions in susceptible individuals called “gluten-related disorders (GRDs)”, which include celiac disease (CD), wheat allergy (WA), and non-celiac gluten sensitivity (NCGS). Currently removing gluten from the diet is the only effective treatment for mentioned GRDs and studies for the appropriate and alternative therapeutic approaches are ongoing. Accordingly, several genetic studies have focused on breeding wheat with low immunological properties through gene editing methods. The present review considers genetic characteristics of gluten protein components, focusing on their role in the incidence of gluten-related diseases, and genetic modifications conducted to produce wheat with less immunological properties.

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“…Because a-gliadin genes are located in tandem arrays, genome editing targeting conserved regions within these genes simultaneously may result in partial or complete deletion of neighboring gliadin genes copies, and evidence for that effect was also obtained (Jouanin et al 2020a). The low-gluten wheat lines obtained in these studies provide a basis for subsequent editing studies to further reduce the epitope load in these wheat lines and to ultimately generate hypoimmunogenic-gluten wheat varieties (Jouanin et al 2020b).…”

Section: Genome Editing and Polyploid Cropsmentioning

confidence: 99%

Genome editing of polyploid crops: prospects, achievements and bottlenecks

2021

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Plant breeding aims to develop improved crop varieties. Many crops have a polyploid and often highly heterozygous genome, which may make breeding of polyploid crops a real challenge. The efficiency of traditional breeding based on crossing and selection has been improved by using marker-assisted selection (MAS), and MAS is also being applied in polyploid crops, which helps e.g. for introgression breeding. However, methods such as random mutation breeding are difficult to apply in polyploid crops because there are multiple homoeologous copies (alleles) of each gene. Genome editing technology has revolutionized mutagenesis as it enables precisely selecting targets. The genome editing tool CRISPR/Cas is especially valuable for targeted mutagenesis in polyploids, as all alleles and/or copies of a gene can be targeted at once. Even multiple genes, each with multiple alleles, may be targeted simultaneously. In addition to targeted mutagenesis, targeted replacement of undesirable alleles by desired ones may become a promising application of genome editing for the improvement of polyploid crops, in the near future. Several examples of the application of genome editing for targeted mutagenesis are described here for a range of polyploid crops, and achievements and bottlenecks are highlighted.

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CRISPR/Cas9 Gene Editing of Gluten in Wheat to Reduce Gluten Content and Exposure—Reviewing Methods to Screen for Coeliac Safety

Cited by 67 publications

References 104 publications

Exploring the alpha‐gliadin locus: the 33‐mer peptide with six overlapping coeliac disease epitopes in Triticum aestivum is derived from a subgroup of Aegilops tauschii

Exploring the alpha‐gliadin locus: the 33‐mer peptide with six overlapping coeliac disease epitopes in Triticum aestivum is derived from a subgroup of Aegilops tauschii

The Gluten Gene: Unlocking the Understanding of Gluten Sensitivity and Intolerance

Genome editing of polyploid crops: prospects, achievements and bottlenecks

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