Related Experiment Video
Updated: Apr 2, 2026
11:27
Efficient Production and Identification of CRISPR/Cas9-generated Gene Knockouts in the Model System Danio rerio
Published on: August 28, 2018
23.3K
Generating adamts13-/- Zebrafish via CRISPR/Cas9 Gene Editing.
Liang Zheng1,2, Zhijian Wu3, X Long Zheng3,4
1Department of Pathology and Laboratory Medicine, The University of Kansas Medical Center, Kansas City, KS, USA. lzheng@kumc.edu.
Methods in Molecular Biology (Clifton, N.J.)
|March 31, 2026
Summary
This study details creating adamts13 knockout zebrafish using CRISPR/Cas9 gene editing. This method establishes a robust framework for studying thrombotic diseases and ADAMTS13 function in vivo.
Area of Science:
- Genetics and Genomics
- Developmental Biology
- Hematology
Background:
- Zebrafish (Danio rerio) are valuable vertebrate models for studying hematologic and thrombotic diseases.
- They possess genetic tractability and conserved hemostatic pathways with humans.
- ADAMTS13 plays a critical role in regulating von Willebrand factor, and its deficiency is linked to thrombotic microangiopathies.
Purpose of the Study:
- To describe a detailed workflow for generating adamts13 knockout zebrafish.
- To establish a robust framework for creating zebrafish loss-of-function models for studying thrombotic diseases.
- To enable mechanistic studies of ADAMTS13 function in vivo.
Main Methods:
- CRISPR/Cas9-mediated genome editing was employed to generate adamts13 knockout zebrafish.
- Protocols included guide RNA design and synthesis, Cas9 mRNA production, and microinjection into one-cell stage embryos.
- Screening founder fish, genotyping, and establishing stable mutant lines through outcrossing were performed to ensure reproducibility and minimize off-target effects.
Main Results:
- A detailed workflow for generating adamts13 knockout zebrafish was successfully established.
- The procedures allow for the creation of stable mutant lines with minimized potential off-target effects.
- The developed framework enables in vivo studies of ADAMTS13 function.
Conclusions:
- The described procedures provide a robust framework for creating zebrafish loss-of-function models.
- This approach facilitates the study of ADAMTS13 function and thrombotic disease pathophysiology.
- The methodology is adaptable for knocking out other genes or introducing specific mutations in zebrafish.
