The Generation of Zebrafish Mariner Model Using the CRISPR/Cas9 System.

Autor: Zou B; Department of Otolaryngology, University of Miami Miller School of Medicine, Miami, Florida.; Department of Biology, University of Miami, Miami, Florida., Desmidt AA; Department of Biology, University of Miami, Miami, Florida., Mittal R; Department of Otolaryngology, University of Miami Miller School of Medicine, Miami, Florida., Yan D; Department of Otolaryngology, University of Miami Miller School of Medicine, Miami, Florida., Richmond M; Department of Biology, University of Miami, Miami, Florida., Tekin M; Dr. John T. Macdonald Foundation Department of Human Genetics and John P. Hussman Institute for Human Genomics, University of Miami Miller School of Medicine, Miami, Florida., Liu XZ; Department of Otolaryngology, University of Miami Miller School of Medicine, Miami, Florida.; Department of Otolaryngology, Xiangya Hospital, Central South University, Changsha, Hunan, China., Lu Z; Department of Biology, University of Miami, Miami, Florida.; Neuroscience Program, University of Miami, Miami, Florida.; International Center for Marine Studies, Shanghai Ocean University, Shanghai, China.
Jazyk: angličtina
Zdroj: Anatomical record (Hoboken, N.J. : 2007) [Anat Rec (Hoboken)] 2020 Mar; Vol. 303 (3), pp. 556-562. Date of Electronic Publication: 2019 Jul 18.
DOI: 10.1002/ar.24221
Abstrakt: Targeted genome editing mediated by clustered, regularly interspaced, short palindromic repeat (CRISPR)/CRISPR-associated nuclease 9 (Cas9) technology has emerged as a powerful tool for gene function studies and has great potential for gene therapy. Although CRISPR/Cas9 has been widely used in many research fields, only a few successful zebrafish models have been established using this technology in hearing research. In this study, we successfully created zebrafish mariner mutants by targeting the motor head domain of Myo7aa using CRISPR/Cas9. The CRISPR/Cas9-generated mutants showed unbalanced swimming behavior and disorganized sterocilia of inner ear hair cells, which resemble the phenotype of the zebrafish mariner mutants. In addition, we found that CRISPR/Cas9-generated mutants have reduced number of stereociliary bundles of inner ear hair cells and have significant hearing loss. Furthermore, phenotypic analysis was performed on F0 larvae within the first week post fertilization, which dramatically shortens data collection period. Therefore, results of this study showed that CRISPR/Cas9 is a quick and effective method to generate zebrafish mutants as a model for studying human genetic deafness. Anat Rec, 303:556-562, 2020. © 2019 American Association for Anatomy.
(© 2019 Wiley Periodicals, Inc.)
Databáze: MEDLINE
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