Using two guide RNAs with CRISPR/Cas9 to efficiently delete large genes in rabbits

A large deletion of 105 kb in the TYR gene was generated in rabbits using a dual sgRNA-directed CRISPR/Cas9 system.

• The created TYR knockout rabbits exhibited typical symptoms of albinism.
• Significantly decreased expression of the TYR gene was observed in the knockout rabbits.
• The same genotype and albinism phenotype were transmitted to the F1 generation.
• The dual sgRNA-directed CRISPR/Cas9 system enabled mono and biallelic large deletions.
• This approach produced no mosaic mutations or off-target effects.

Simplified