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

Bing Zou, Alexandra A. Desmidt, Rahul Mittal, Denise Yan, Micheal Richmond, Mustafa Tekin, Xuezhong Liu, Zhongmin Lu

Research output: Contribution to journalArticle

Abstract

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, 2019.

Original languageEnglish (US)
JournalAnatomical Record
DOIs
StateAccepted/In press - Jan 1 2019

Fingerprint

Clustered Regularly Interspaced Short Palindromic Repeats
nucleases
Zebrafish
Danio rerio
mutants
hearing
hair
Inner Auditory Hair Cells
hairs
ears
swimming behavior
gene
deafness
gene therapy
targeting
Inner Ear
phenotype
genome
mutant
larva

Keywords

  • CRISPR/Cas9
  • hearing loss
  • mariner
  • myo7aa
  • zebrafish

ASJC Scopus subject areas

  • Anatomy
  • Biotechnology
  • Histology
  • Ecology, Evolution, Behavior and Systematics

Cite this

The Generation of Zebrafish Mariner Model Using the CRISPR/Cas9 System. / Zou, Bing; Desmidt, Alexandra A.; Mittal, Rahul; Yan, Denise; Richmond, Micheal; Tekin, Mustafa; Liu, Xuezhong; Lu, Zhongmin.

In: Anatomical Record, 01.01.2019.

Research output: Contribution to journalArticle

@article{9c8762dfe0a44e43b43a98b7ba68e8e9,
title = "The Generation of Zebrafish Mariner Model Using the CRISPR/Cas9 System",
abstract = "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, 2019.",
keywords = "CRISPR/Cas9, hearing loss, mariner, myo7aa, zebrafish",
author = "Bing Zou and Desmidt, {Alexandra A.} and Rahul Mittal and Denise Yan and Micheal Richmond and Mustafa Tekin and Xuezhong Liu and Zhongmin Lu",
year = "2019",
month = "1",
day = "1",
doi = "10.1002/ar.24221",
language = "English (US)",
journal = "Anatomical Record",
issn = "1932-8486",
publisher = "John Wiley and Sons Inc.",

}

TY - JOUR

T1 - The Generation of Zebrafish Mariner Model Using the CRISPR/Cas9 System

AU - Zou, Bing

AU - Desmidt, Alexandra A.

AU - Mittal, Rahul

AU - Yan, Denise

AU - Richmond, Micheal

AU - Tekin, Mustafa

AU - Liu, Xuezhong

AU - Lu, Zhongmin

PY - 2019/1/1

Y1 - 2019/1/1

N2 - 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, 2019.

AB - 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, 2019.

KW - CRISPR/Cas9

KW - hearing loss

KW - mariner

KW - myo7aa

KW - zebrafish

UR - http://www.scopus.com/inward/record.url?scp=85069924617&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85069924617&partnerID=8YFLogxK

U2 - 10.1002/ar.24221

DO - 10.1002/ar.24221

M3 - Article

C2 - 31260171

AN - SCOPUS:85069924617

JO - Anatomical Record

JF - Anatomical Record

SN - 1932-8486

ER -