ArticleMicrobial biotechnology2026
Exploiting the Endogenous Type II-A CRISPR-Cas System for Functional Engineering of Probiotic Lacticaseibacillus rhamnosus GG.
Article in Microbial biotechnology, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed.
- Exploring the emerging role of CRISPR-Cas systems in probiotic development.Engineering microbiology · 2026Review
- Enhancing Probiotic Performance in Food Matrices: From Strain Selection to Bioengineering.Journal of food science · 2026Review
- Nutrient control enables metabolic reconstruction ofbioRxiv : the preprint server for biology · 2026Article
- Genetic Engineering and Encapsulation Strategies forFoods (Basel, Switzerland) · 2026Review
- Exploiting the Endogenous Type II-A CRISPR-Cas System for Functional Engineering of Probiotic Lacticaseibacillus rhamnosus GG.Microbial biotechnology · 2026Article
- Exploring the probiotic landscape in understanding postbiotics from indigenous bacteria isolated from the stool samples of a tribal population at Mulluvadi village, Tamil Nadu, India.Frontiers in pharmacology · 2026Article
Corrections and comments
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Authors and funding
3 authors.
Funding
Abstract
Lacticaseibacillus rhamnosus GG (LGG) is one of the most extensively studied probiotic strains, widely used in food and health applications. However, the absence of efficient, precise genome editing methods has limited its broader potential and functional versatility. Here, we present an endogenous type II-A CRISPR-Cas genome editing workflow for LGG designed for functional strain construction. Using a plasmid interference assay together with single-nucleotide substitutions, we confirm the precise PAM requirement as 5'-NGAAA-3'. We pair a synthetic sgRNA cassette with homology-directed repair donors to enable targeted deletions and insertions across multiple loci, achieving modest but practically relevant editing efficiencies (11.1-25.0% of recovered transformants) that support routine strain construction. Using this optimised and precise genome engineering method, we generated a β-glucuronidase (GUS)-expressing LGG strain for robust strain tracking within complex microbial communities. This work removes barriers to LGG engineering, expands the probiotic CRISPR toolkit, and provides broadly applicable strategies for designing next-generation probiotics with applications in food biotechnology and microbial therapeutics.
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Registered trials
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