Improving Bacillus subtilis as Biological Chassis Performance by the CRISPR Genetic Toolkit

ACS Synthetic Biology, Journal Year: 2025, Volume and Issue: unknown

Published: March 4, 2025

Bacillus subtilis is the model Gram-positive and industrial chassis bacterium; it has blossomed as a robust promising host for enzyme, biochemical, or bioflocculant production. However, synthetic biology metabolic engineering technologies of B. have lagged behind most widely used Saccharomyces cerevisiae Escherichia coli. CRISPR (an acronym clustered regularly interspaced short palindromic repeats) enables efficient, site-specific, programmable DNA cleavage, which revolutionized manner genome editing. In 2016, technology was first introduced into been intensely upgraded since then. this Review, we discuss recently developed key additions to toolkit design in with gene editing, transcriptional regulation, enzyme modulation. Second, advances efficient biochemicals proteins are discussed. Finally, conclude perspectives on challenges opportunities CRISPR-based biotechnology subtilis, wishing that can be comparable traditional microorganisms such E. coli S. someday soon.

Language: Английский

CRISPR-Based Regulation for High-Throughput Screening

ACS Synthetic Biology, Journal Year: 2025, Volume and Issue: unknown

Published: May 22, 2025

CRISPR technology has revolutionized genome editing by enabling precise, permanent modifications to genetic material. To circumvent the irreversible alterations associated with traditional methods and facilitate research on both essential nonessential genes, interference or inhibition (CRISPRi) activation (CRISPRa) were developed. The gene-silencing approach leverages an inactivated Cas effector protein paired guide RNA obstruct transcription initiation elongation, while gene-activation exploits programmability of activate gene expression. Recent advances in CRISPRi technology, combination other technologies (e.g., biosensing, sequencing), have significantly expanded its applications, allowing for genome-wide high-throughput screening (HTS) identify determinants phenotypes. These strategies been applied biomedicine, industry, basic research. This review explores regulation mechanisms, offers overview workflow CRISPR-based screens, highlights superior suitability HTS across biomedical industrial applications. Finally, we discuss limitations current CRISPRi/a envision future directions CRISPR-mediated research, considering potential broader application diverse fields.

Language: Английский

Intelligent biomanufacturing of water-soluble vitamins

Trends in biotechnology, Journal Year: 2025, Volume and Issue: unknown

Published: May 1, 2025

Language: Английский