Targeted mutagenesis in mice via an engineered AsCas12f1 system

Area of Science:

• Molecular Biology
• Gene Editing Technologies
• Biotechnology

Background:

• SpCas9 and AsCas12a are standard genome editing tools but are limited by their large size.
• AsCas12f1 is a smaller alternative, but its application is hindered by low and variable editing efficiency.

Purpose of the Study:

• To enhance the editing efficiency of the AsCas12f1 system.
• To explore the therapeutic potential of the engineered AsCas12f1 system.
• To generate novel mouse models for disease research.

Main Methods:

• Engineering of the AsCas12f1 single-guide RNA (sgRNA).
• Testing the engineered CRISPR-AsCas12f1 system in human cells and mouse embryos.
• Generation of stable mouse mutant disease models.

Main Results:

• Engineered AsCas12f1 sgRNA significantly improved editing efficiency.
• Successful generation of three stable mouse mutant disease models.
• Demonstrated broad applicability and enhanced performance of the mini CRISPR system.

Conclusions:

• The engineered AsCas12f1 system expands the mini CRISPR toolbox.
• This enhanced system shows significant promise for therapeutic applications.
• The study provides a valuable tool for genome editing and disease modeling.