Activating natural product synthesis using CRISPR interference and activation systems in Streptomyces

Area of Science:

• Microbiology
• Synthetic Biology
• Drug Discovery

Background:

• Antibiotic resistance poses a significant global health threat, necessitating new antibiotic discovery.
• Streptomyces bacteria are a key source of natural product antibiotics, but many are difficult to produce.
• Silent biosynthetic gene clusters (BGCs) limit the discovery of novel compounds from Streptomyces.

Purpose of the Study:

• To develop novel synthetic gene regulators for activating silent BGCs in Streptomyces.
• To engineer CRISPR interference (CRISPRi) and CRISPR activation (CRISPRa) systems for Streptomyces.
• To facilitate the discovery of new antibiotics by activating previously inaccessible BGCs.

Main Methods:

• Refinement of CRISPR interference (CRISPRi) for gene repression in Streptomyces.
• Development of CRISPR activation (CRISPRa) for gene activation in Streptomyces.
• Application of CRISPR tools to perturb regulatory networks and activate silent BGCs.

Main Results:

• Demonstration of programmable gene repression and activation in Streptomyces using CRISPR systems.
• Successful activation of a silent BGC by manipulating its endogenous regulatory network.
• Expansion of the synthetic regulatory toolbox for Streptomyces.

Conclusions:

• The developed CRISPR-based tools enhance the activation of silent BGCs in Streptomyces.
• This approach facilitates the discovery of novel natural products and antibiotics.
• Programmable activation of BGCs is a powerful strategy for addressing antimicrobial resistance.