Xingzhu Feng

Hainan Institute of Biotechnology, Haikou, 570206, Hainan, China
Author    Correspondence author
Legume Genomics and Genetics, 2025, Vol. 16, No. 2   
Received: 30 Jan., 2025    Accepted: 15 Mar., 2025    Published: 05 Apr., 2025

This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Trypsin inhibitors (TIs) in soybean are known to have antinutritional effects, reducing protein digestibility and limiting the nutritional value of soy products and animal feeds. To address this long-standing challenge, genome editing tools such as CRISPR/Cas9 have emerged as promising strategies for precisely eliminating undesirable traits such as TIs. This study explores the application of CRISPR/Cas9 to targetedly ablate trypsin inhibitor genes in soybean, specifically those encoding Kunitz and Bowman-Birk inhibitors. We discuss the biological functions and limitations of these inhibitors, outline the mechanisms and recent technical improvements of CRISPR/Cas9, and detail methods for identifying TI gene targets using transcriptomic and proteomic analyses. We also review guide RNA design, translational techniques, and gene editing validation. Functional assessments demonstrated that knockout lines exhibited reduced TI activity, improved protein digestibility, and improved nutritional status, with minimal adverse effects on agronomic traits. A case study demonstrating the successful ablation of the Kunitz trypsin inhibitor gene further demonstrates the utility of this approach. We also explore biosafety concerns, regulatory frameworks, and public perception issues surrounding genome-edited crops. Ultimately, this study highlights the transformative potential of CRISPR/Cas9 for improving the nutritional quality of soybeans and supports future efforts to integrate genome editing into breeding programs to develop high-protein, low-antinutrient varieties.

CRISPR/Cas9; Trypsin Inhibitors; Soybean; Genome Editing; Nutritional Enhancement