Related Experiment Video
Updated: Sep 8, 2025
09:40
Generation of Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9
Published on: January 3, 2015
95.8K
Generation of a Human Deafness Sheep Model Using the CRISPR/Cas System
Martina Crispo1, Vanessa Chenouard2, Pedro Dos Santos-Neto3
1Laboratory Animal Biotechnology Unit, Institut Pasteur de Montevideo, Montevideo, Uruguay.
Methods in Molecular Biology (Clifton, N.J.)
|June 13, 2022
Summary
Researchers created a sheep model for human deafness using CRISPR gene editing. This otoferlin knock-in model will aid in developing new therapies for auditory neuropathy spectrum disorder.
Area of Science:
- Genetics and Genomics
- Animal Models
- Biotechnology
Background:
- The CRISPR/Cas9 system enables genome editing for creating human disease models in animals.
- Sheep offer advantages as medium/large animal models for studying human diseases.
- Mutations in the otoferlin (OTOF) gene cause nonsyndromic recessive auditory neuropathy spectrum disorder.
Purpose of the Study:
- To establish a sheep model for human deafness by editing the otoferlin gene.
- To develop a protocol for generating gene-edited sheep using CRISPR-Cas9 and homology-directed repair.
- To create a valuable preclinical model for testing therapies targeting OTOF mutations.
Main Methods:
- CRISPR-assisted single-stranded oligodeoxynucleotide-mediated Homology-Directed Repair (HDR).
- Direct cytoplasmic microinjection into in vitro produced sheep zygotes.
- Generation of otoferlin knock-in (KI) sheep.
Main Results:
- Successfully generated an otoferlin edited sheep model.
- 17.8% of lambs exhibited indel mutations in the OTOF gene.
- 61.5% of edited lambs carried knock-in mutations via HDR.
Conclusions:
- The developed protocol provides a foundation for creating otoferlin knock-in sheep.
- This model serves as a platform for investigating therapies for OTOF-related human deafness.
- Advances in genome editing facilitate the creation of relevant animal models for human genetic disorders.
