Working Paper Review Version 2 This version is not peer-reviewed

Current Status and Perspectives on the Application of CRISPR/Cas9 Gene-Editing System to Develop a Low-Gluten, Non-Transgenic Wheat Variety

Version 1 : Received: 19 February 2020 / Approved: 21 February 2020 / Online: 21 February 2020 (02:14:36 CET)
Version 2 : Received: 28 September 2021 / Approved: 29 September 2021 / Online: 29 September 2021 (15:39:12 CEST)

A peer-reviewed article of this Preprint also exists.

Journal reference: Foods 2021, 10
DOI: 10.3390/foods10102351


Wheat gluten contains epitopes that trigger celiac disease (CD). A life-long strict gluten-free diet is the only treatment accepted for CD. However, very low-gluten wheat may provide an alternative treatment to CD. Conventional plant breeding methods are not sufficient to produce celiac-safe wheat. RNA interference technology, to some extent, succeeded in the development of safer wheat varieties. However, these varieties had multiple challenges in their implementation. Clustered Regularly Interspaced Short Palindromic Repeats-associated nuclease 9 (CRISPR/Cas9) is a versatile gene-editing tool that has the ability to edit the immunogenic gluten genes. So far, only a few studies have applied CRISPR/Cas9 to modify the wheat genome. In this article, we reviewed published literature that applied CRISPR/Cas9 in wheat genome editing to investigate the current status of the CRISPR/Cas9 system to produce a low-immunogenic wheat variety. We found that in recent years, the CRISPR/Cas9 system has been continuously improved to edit the complex hexaploid wheat genome. Although some reduced immunogenic wheat varieties have been reported, CRISPR/Cas9 has still not been fully explored in editing the wheat genome. We conclude that further studies are required to apply the CRISPR/Cas9 gene-editing system efficiently for the development of celiac-safe wheat variety and to establish it as a “tool to celiac safe wheat.”


CRISPR/Cas9; celiac disease; wheat; sgRNA; gluten; low-immunogenic wheat



Comments (1)

Comment 1
Received: 29 September 2021
Commenter: Anil K. Verma
Commenter's Conflict of Interests: Author
Comment: The article has been revised. Multiple changes have been done. 
The major changes are:
* Study title has been revised
*An author has been  replaced with another
*RNAi method has been trimmed
*A figure showing a comparison between RNAi and CRISPR has been added
*13 latest studies have been added in Table 1 (in total 23 studies now)
*CRISPR methodology figure has been revised
* English grammar, punctuations have been rechecked carefully 

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