Advances in multiplex precision genome editing in eukaryotic and prokaryotic systems

Area of Science:

• Molecular Biology
• Biotechnology
• Genomics

Background:

• Multiplex genome editing (MGE) is crucial for engineering complex traits.
• Traditional CRISPR-Cas systems using DNA double-strand breaks (DSBs) face precision and scalability limits for MGE.
• Emerging DSB-free genome editing technologies offer enhanced accuracy for multiplexed genomic manipulation.

Purpose of the Study:

• To review recent advances in precise Multiplex Genome Editing (MGE) strategies.
• To highlight key engineering principles for scalable MGE.
• To discuss the applications of precise MGE across various biological systems.

Main Methods:

• Focus on base editing, prime editing, and related genome rewriting platforms.
• Analysis of engineering principles: editing platform choice, edit size, and guide RNA architecture.
• Review of applications in mammalian, plant, fungal, and bacterial systems.

Main Results:

• DSB-free genome editing technologies significantly improve precision and feasibility of MGE.
• Key factors like editing platform, edit size, and guide RNA design are critical for successful scalable multiplexing.
• Precise MGE strategies are applicable across a wide range of organisms.

Conclusions:

• Emerging precise MGE technologies overcome limitations of traditional methods.
• These advancements enable versatile, precise control over multiple genes.
• Precise MGE holds significant potential for biotechnology and agriculture.