Phenotypes of mutant cell lines and determination of target metabolites at different culture durations. (A) Phenotypes and microscopic observations of CHS2 mutant cell lines. Determination of (B) flavonoids, (C) anthocyanins, (D) proanthocyanindins, (E) resveratrol, and (F) piceid.
Resveratrol has garnered attention for its potential anti-aging, anti-inflammatory, and heart-health benefits. However, natural sources such as grapes and peanuts yield only small amounts, and current production methods face significant limitations. Extracting resveratrol from plants is inefficient, chemical synthesis often leads to impurities, and microbial fermentation struggles to properly fold proteins. In plants, resveratrol competes with flavonoids, another class of compounds. Because the flavonoid pathway typically dominates, resveratrol production remains minimal. These challenges have hindered the widespread availability of resveratrol for both research and commercial use. To overcome these obstacles, researchers sought a way to redirect the plant’s metabolic pathways to increase resveratrol yields.
On October 9, 2024, scientists from Fujian Academy of Agricultural Sciences and Shanghai Jiao Tong University published a pivotal in , where they used CRISPR/Cas9 to edit grape cells (Vitis davidii). By targeting the CHS2 gene, which is responsible for flavonoid production, they reduced the competition for biochemical precursors, resulting in a more than 400% increase in resveratrol production. This study provides the first evidence that CRISPR can be used to shift metabolic pathways to favor valuable compounds like resveratrol, while also establishing grape cell cultures as a promising production platform, avoiding the issues of microbial systems and whole plant extraction.
The research team used CRISPR to create two mutant grape cell lines by editing the CHS2 gene. This gene plays a pivotal role in flavonoid production. Their detailed analysis confirmed that the genetic edits successfully redirected the cells' metabolic resources towards resveratrol synthesis. The most effective mutant line (MT1) produced 4.22 μg of resveratrol per gram of fresh cells, a fourfold increase compared to unedited cells. The derivative of resveratrol, piceid, saw an even greater increase of 5.3-fold. Genetic analysis confirmed that the CHS2 gene was almost completely knocked out (99.87% mutation rate), leading to a significant downregulation of flavonoid pathway genes, while resveratrol production genes became more active. The researchers also verified that the edits were precise, with no unintended effects on other genes. Remarkably, the position of the genetic edit was crucial — modifications closer to the start of the gene (the 5' region) led to more complete disruption of flavonoid production. Over a 40-day period, cell cultures maintained stable resveratrol production, suggesting this method could be scaled up for industrial use.
“These results provide a roadmap for using precision gene editing to turn plant cells into efficient factories for health-promoting compounds,” said corresponding author Dr. Chengchun Lai. “By understanding and manipulating the competition between metabolic pathways, we've developed a more sustainable way to produce resveratrol that avoids the pitfalls of current methods. This approach could be adapted to enhance the production of other rare but valuable plant molecules.”
This breakthrough has the potential to change the landscape of resveratrol production for multiple industries. Nutraceutical and pharmaceutical companies may adopt this cell-based method to produce purer and more sustainable resveratrol for use in supplements and medicines. The food industry could more economically incorporate resveratrol into functional foods. Agriculturally, this technique may offer new methods for breeding crops with enhanced beneficial compounds without relying on traditional genetic modification. Moreover, the principles developed in this study could be applied to the production of other valuable plant-derived compounds that are currently difficult to obtain. Future research will focus on optimizing culture conditions to increase yields further and explore the scalability of this method for commercial production. As demand for plant-based bioactive compounds continues to rise, biotechnological advancements like these will play a key role in meeting this need.
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Funding information
This work was founded by National Natural Science Foundation of China (32302284), Natural Science Foundation of Fujian Province, China (2021 J05091), High Quality Development “5511” Collaborative Innovation Project between Fujian and Chinese Academy of Agricultural Sciences (XTCXGC2021014), and Fujian Provincial Department of Science and Technology of Special Public-funded Projects (2021R1032009).
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