Omega-based mini prime- and base-editing systems in Escherichia coli

Area of Science:

• Molecular Biology
• Biotechnology
• Synthetic Biology

Background:

• Prime editing (PE) and base editing (BE) are advanced gene-editing technologies.
• Their large size limits efficient delivery and broad use.
• Developing smaller, efficient gene editing tools is crucial.

Purpose of the Study:

• To engineer and evolve compact obligate mobile element-guided activity (OMEGA)-based PE and BE systems.
• To improve editing efficiency, specificity, and versatility.
• To address delivery and application bottlenecks of current gene editing tools.

Main Methods:

• Engineering and evolution of OMEGA-based prime editing (OMEGA-PE) and adenine base editing systems.
• Testing editing efficiency and specificity in Escherichia coli.
• Assessing compatibility with high-throughput screening platforms.

Main Results:

• OMEGA-PE and OMEGA-adenine base editor demonstrated superior editing efficiency and versatility in E. coli compared to existing tools.
• OMEGA-PE achieved rapid, programmable targeted mutagenesis with high specificity and efficiency.
• The systems showed excellent compatibility with high-throughput screening and enhanced cofactor regeneration and protein translation.

Conclusions:

• Compact OMEGA-based gene editing systems overcome size limitations of traditional PE and BE.
• These tools offer enhanced performance for gene editing, directed evolution, and metabolic engineering.
• OMEGA-based systems provide new possibilities for basic research and industrial biotechnology.