Genetic Modification of Mice Using Prime Editing.

Area of Science:

• Molecular Biology
• Genetics
• Animal Models

Background:

• Traditional mouse genetic modification is labor-intensive, requiring extensive steps like construct design, stem cell manipulation, and chimera breeding.
• These conventional methods are time-consuming (often over a year), costly, and lack guaranteed success, necessitating specialized facilities or vendors.
• The advent of RNA-guided CRISPR systems has simplified gene editing, enabling efficient modifications in mice and other challenging animal models.

Purpose of the Study:

• To present a streamlined protocol for generating genetically modified mice.
• To introduce the prime editing platform as a simplified method for mouse genetic engineering.
• To detail a simplified approach for creating gene modifications in mice.

Main Methods:

• Design, cloning, and synthesis of engineered prime editing guide RNA (pegRNA).
• Microinjection of prime editing (PE2) components into mouse zygotes.
• Genotyping of founder mice and subsequent breeding for germline transmission.

Main Results:

• The prime editing platform facilitates the generation of gene modifications (substitutions, insertions, deletions) in mice with high efficiency.
• This method simplifies the complex process of creating genetically altered mice.
• Successful germline transmission was achieved through the described protocols.

Conclusions:

• Prime editing provides a significantly simplified and efficient alternative to traditional methods for generating genetically modified mice.
• This protocol enables high-success-rate gene editing in mice, reducing project timelines and complexity.
• The described approach democratizes the generation of gene-edited mouse models.