Practical Approaches for Knock-Out Gene Editing in Pigs.

Area of Science:

• Genetics and Genomics
• Biomedical Research
• Animal Models

Background:

• Pigs are valuable for meat production and as models for human diseases due to physiological similarities.
• Previous limitations in genetic modification hindered widespread use of pigs in research.
• The advent of programmable nucleases, particularly CRISPR-Cas9, has revolutionized the ability to modify the porcine genome.

Purpose of the Study:

• To review strategies for gene editing in pigs using the CRISPR-Cas9 system.
• To discuss methods for delivering gene editing tools into porcine embryos.
• To explore approaches for producing homogeneous, biallelic gene-edited pigs.

Main Methods:

• Summarizing various CRISPR-Cas9 based gene editing strategies for pigs.
• Detailing genotyping techniques for assessing gene edits.
• Reviewing different delivery methods for introducing CRISPR-Cas9 components into zygotes or cells.

Main Results:

• CRISPR-Cas9 allows for unprecedented precision in modifying the porcine genome.
• Successful gene editing requires efficient delivery into early-stage embryos or edited somatic cells for nuclear transfer.
• Strategies exist to achieve biallelic gene editing, leading to uniformly modified animals.

Conclusions:

• CRISPR-Cas9 technology overcomes previous hurdles in porcine genetic modification.
• Effective strategies and delivery methods are crucial for generating precisely edited pigs.
• This technology significantly enhances the utility of pigs as models for human disease research and biomedical applications.