Utilization of Native CRISPR-Cas9 System for Expression of Glucagon-like Peptide-1 in Lacticaseibacillus paracasei

Zheng, Mumin; Zhang, Shuwen; Wang, Yunna; Xie, Ning; Wang, Xiaodan; Lv, Jiaping; Pang, Xiaoyang; Li, Xu

doi:10.3390/foods14101785

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Open AccessArticle

Utilization of Native CRISPR-Cas9 System for Expression of Glucagon-like Peptide-1 in Lacticaseibacillus paracasei

by

Mumin Zheng

,

Shuwen Zhang

,

Yunna Wang

,

Ning Xie

,

Xiaodan Wang

,

Jiaping Lv

,

Xiaoyang Pang

^*

and

Xu Li

^*

Key Laboratory of Agro-Products Quality and Safety Control in Storage and Transport Process, Ministry of Agriculture and Rural Affairs, Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences, Beijing 100193, China

^*

Authors to whom correspondence should be addressed.

Foods 2025, 14(10), 1785; https://doi.org/10.3390/foods14101785 (registering DOI)

Submission received: 6 April 2025 / Revised: 3 May 2025 / Accepted: 14 May 2025 / Published: 17 May 2025

(This article belongs to the Section Food Microbiology)

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Abstract

Type 2 diabetes is one of the main causes of cardiovascular diseases, kidney diseases, and visual impairments, posing a global healthcare challenge. The current treatment of this disease, involving glucagon-like peptide-1 (GLP-1), is faced with problems such as frequent injections and plasmid instability. In this study, we used the native clustered regularly interspaced short palindromic repeats-CRISPR-associated protein 9 (CRISPR-Cas9) system of Lacticaseibacillus paracasei to develop a novel, genetically stable, and orally administrable strain expressing human GLP-1. Integration and subsequent expression of glp-1 gene were confirmed by genomic sequencing, qPCR, and Nano LC-MS. The engineered strain demonstrated stable genomic integration and sustained high-level expression of GLP-1 over multiple generations. This innovative approach provides a promising strategy for the oral delivery of therapeutic peptides, potentially enhancing patient compliance and improving the treatment of diabetes and other chronic diseases requiring peptide-based therapies.

Keywords: Lacticaseibacillus paracasei; native CRISPR-Cas9; glucagon-like peptide-1 (GLP-1); genome editing; heterologous expression

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MDPI and ACS Style

Zheng, M.; Zhang, S.; Wang, Y.; Xie, N.; Wang, X.; Lv, J.; Pang, X.; Li, X. Utilization of Native CRISPR-Cas9 System for Expression of Glucagon-like Peptide-1 in Lacticaseibacillus paracasei. Foods 2025, 14, 1785. https://doi.org/10.3390/foods14101785

AMA Style

Zheng M, Zhang S, Wang Y, Xie N, Wang X, Lv J, Pang X, Li X. Utilization of Native CRISPR-Cas9 System for Expression of Glucagon-like Peptide-1 in Lacticaseibacillus paracasei. Foods. 2025; 14(10):1785. https://doi.org/10.3390/foods14101785

Chicago/Turabian Style

Zheng, Mumin, Shuwen Zhang, Yunna Wang, Ning Xie, Xiaodan Wang, Jiaping Lv, Xiaoyang Pang, and Xu Li. 2025. "Utilization of Native CRISPR-Cas9 System for Expression of Glucagon-like Peptide-1 in Lacticaseibacillus paracasei" Foods 14, no. 10: 1785. https://doi.org/10.3390/foods14101785

APA Style

Zheng, M., Zhang, S., Wang, Y., Xie, N., Wang, X., Lv, J., Pang, X., & Li, X. (2025). Utilization of Native CRISPR-Cas9 System for Expression of Glucagon-like Peptide-1 in Lacticaseibacillus paracasei. Foods, 14(10), 1785. https://doi.org/10.3390/foods14101785

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Utilization of Native CRISPR-Cas9 System for Expression of Glucagon-like Peptide-1 in Lacticaseibacillus paracasei

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