Efficient generation of CLPG1-edited rabbits using the CRISPR/Cas9 system.

Area of Science:

• Genetics
• Animal Models
• Molecular Biology

Background:

• The callipyge (CLPG) phenotype is a muscular hypertrophy syndrome in sheep.
• Its complex mechanisms are not fully understood, partly due to a lack of suitable animal models with CLPG mutations.

Purpose of the Study:

• To confirm the conservation of the CLPG1 locus, including the 12-bp CLPG1 motif, in mammals.
• To generate CLPG1-edited rabbits as a novel animal model for studying the CLPG phenotype.

Main Methods:

• Utilized the CRISPR/Cas9 gene-editing system combined with cytoplasm injection technology.
• Generated seven CLPG1-edited rabbits with high efficiency.
• Screened nine potential off-target sites to assess editing precision.

Main Results:

• Confirmed conservation of the CLPG1 locus and motif in rabbits.
• Successfully generated CLPG1-edited rabbits with 100% efficiency, though newborns exhibited mosaicism.
• Detected no off-target effects at the nine screened sites, indicating precise gene editing.

Conclusions:

• CLPG1-edited rabbits were efficiently and precisely generated using CRISPR/Cas9.
• These rabbits represent a valuable new animal model for investigating the intricate mechanisms of the callipyge phenotype.