Where genetic accessibility can be established, the integration of CRISPR technology with synthetic biology may enable more precise gene regulation and could support the development of next-generation engineered Bifidobacterium-based platforms.
Abstract
Bifidobacterium plays a vital role in gut health, immunity, and metabolism, yet natural strains show limited native functional capabilities and genetic manipulability. Recent advances in genome editing, especially CRISPR-based systems, have provided novel approaches for engineering Bifidobacterium, though strain-specific variability and restriction-modification barriers remain substantial constraints. This review outlines the evolution from traditional homologous recombination to CRISPR/Cas technologies, highlighting applications of endogenous and exogenous systems, including CRISPRi/a and base editing. In preclinical and food biotechnology contexts, genetically engineered strains have shown potential as targeted delivery vehicles and functional probiotics. Despite these achievements, challenges such as strain variability, restriction-modification barriers, and biosafety concerns persist. Where genetic accessibility can be established, the integration of CRISPR technology with synthetic biology may enable more precise gene regulation and could support the development of next-generation engineered Bifidobacterium-based platforms.
The principal barriers to CRISPR-Cas9 genome editing of bacteria are discussed, the current strategies for addressing these barriers are evaluated, and emerging approaches aimed at improving the efficiency, reliability and precision in bacterial genome engineering are highlighted.
Abdul Shabbir, Javeria Idrees, A. Khan et al.· Archives of Microbiology· 0 citations
Clustered regularly interspaced short palindromic repeats (CRISPR) and associated (Cas) systems have revolutionized the field of genome engineering by providing versatile, efficient, and programmable tools for precise genetic manipulation. Originally identified as an adaptive immune mechanism in prokaryotes, CRISPR/Cas...
E. Cakiroglu, Serif Senturk· Methods in molecular biology· 0 citations
A validated CRISPR-Cas9 strategy for high extracellular β-galactosidase secretion in the clinically established S. boulardii CNCM I-745 is demonstrated, providing a foundation for future studies where the potential of the developed strains is evaluated in clinically relevant settings.
J. Carvalho, David Sáez Moreno, Carlos E. Costa et al.· Microbiology Research· 0 citations
This review starts by examining the advantages and limitations of various physical methods for delivering Cas9, and highlights key applications of CRISPR systems in epigenetic modifications, and explores the use of CRISPR-Cas9 technology in genome editing, with a particular focus on base editing and prime editing.
Suhail Ahmad Sheikh, W. Shah, Azka Khan et al.· Journal of Population Therap...· 0 citations
This review provides a comprehensive synthesis of a recent advances in CRISPR–Cas technologies and their strategic applications in crop genetics and hybrid breeding, and showcases how these technologies accelerate hybrid breeding by engineering male sterility systems, fixing heterosis, and generating high-throughput mu...
Syed Riaz Ahmed, Jahangir Khan, I. Ijaz et al.· Frontiers in Plant Science· 1 citation
How CRISPR interference, CRISPR activation, base editing, base editing, and prime editing have improved the understanding of disease mechanisms, while creating new opportunities for therapeutic intervention are discussed.
Moon-Soo Kim, Hae Sol Do, H. Jang et al.· Biomolecules· 0 citations
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