Prime editing in mammals: From promise to practicalities

Area of Science:

• Molecular Biology
• Gene Editing Technologies
• Biotechnology

Background:

• Prime editing (PE) is an advanced CRISPR-Cas9 system.
• It fuses reverse transcriptase (RT) to a Cas9 nickase (nCas9).
• Complexed with prime editing guide RNA (pegRNA), PE enables precise genomic modifications without double-strand breaks.

Purpose of the Study:

• To systematically review prime editing performance across diverse models.
• To analyze factors influencing PE efficiency.
• To discuss methods for assessing and troubleshooting PE experiments.

Main Methods:

• Systematic literature review of prime editing studies.
• Analysis of PE efficiency in various cellular and animal models.
• Examination of reporter assays for detecting and selecting edited cells.

Main Results:

• High variability in PE efficiency reported across different studies and models.
• Identification of approaches that improve or impede PE efficiency.
• Discussion of downstream effects and potential causes for variability.

Conclusions:

• Prime editing shows promise for precise gene editing with reduced off-target effects.
• Understanding model-specific performance and troubleshooting strategies is crucial for successful PE applications.
• This review offers insights for researchers utilizing PE in their experiments.