# Evaluation of the Unintended Effects of fad2-1-Gene-Edited Soybean Line AE15 Seeds

## TL;DR

This study used proteomic analysis to assess unintended effects in gene-edited soybean seeds and found fewer changes compared to natural genetic variation.

## Contribution

The study demonstrates that gene editing in soybean seeds causes fewer unintended protein expression changes than natural genetic variation.

## Key findings

- Gene-edited soybean seeds showed fewer differentially expressed proteins compared to wild-type controls.
- Eight commonly differentially expressed proteins were found across all three generations of gene-edited soybean seeds.
- Two proteins—beta-amylase and ER lumen protein-retaining receptor—were consistently upregulated in gene-edited seeds.

## Abstract

A data-independent acquisition (DIA)-based proteomic analysis was performed to evaluate the unintended effects of fad2-1-gene-edited soybean line AE15 seeds. A total of 561, 269, and 227 differentially expressed proteins (DEPs) were identified in seeds from three consecutive generations of AE15 soybean, respectively, and were primarily enriched in Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways related to carbon metabolism, protein processing in the endoplasmic reticulum, and proteasome function. Furthermore, eight commonly differentially expressed proteins (co-DEPs) were detected across all three generations of AE15 soybean seeds, among which two—beta-amylase and endoplasmic reticulum (ER) lumen protein-retaining receptor—exhibited consistently upregulated expression. In the wild-type soybean control groups, 1063, 989, and 671 DEPs were identified across the three comparisons (ZhH302E3/ZhH10, ZhH10/ZhH42, and ZhH42/ZhH302E3), among which 71 co-DEPs were observed. These findings indicate that the protein expression profile alterations resulting from fad2-1 gene editing are considerably less pronounced compared to those caused by natural genetic variation among soybean seeds.