Modeling a cataract disorder in mice with prime editing

Area of Science:

• Genetics and Genomics
• Molecular Biology
• Biotechnology

Background:

• Prime editing is a versatile genome editing technology capable of precise DNA modifications.
• Previous studies have demonstrated prime editing's efficacy in various organisms, but its application for generating deletion-based genetic disease models in mice remained unexplored.

Purpose of the Study:

• To establish a mouse model for a genetic cataract disorder using prime editing.
• To investigate the efficiency and germline transmission of prime editor 3 (PE3)-induced single-base deletions in mice.

Main Methods:

• Microinjection of prime editor 3 (PE3) plasmids into mouse zygotes to induce targeted single-base deletions.
• Phenotypic analysis of resulting mouse offspring to identify cataract development.
• Assessment of germline transmission rates and inheritance patterns of the induced mutation and associated phenotype.

Main Results:

• Successfully generated a mouse model exhibiting a nuclear cataract phenotype due to a targeted single-base deletion.
• Achieved a high G-deletion rate (38.2%) in the targeted gene.
• Demonstrated high rates of germline transmission for the PE3-induced deletion, with phenotypic inheritance of cataract in subsequent generations.

Conclusions:

• Prime genome editing is an effective tool for modeling genetic diseases caused by single nucleotide deletions in mice.
• The developed PE3-based approach offers a powerful strategy for creating precise genetic disease models in vivo.
• This study provides a foundation for utilizing prime editing to generate novel animal models for a wider range of genetic disorders.