Protocol for conditional mutagenesis in zebrafish germ cells using Tol2 transposon and a CRISPR-Cas9-based plasmid system

Yixuan Hu; Yutao Chen; Yu Zhang; Zhiquan Liu; Jianzhen Li

doi:10.1016/j.xpro.2024.103516

Protocol for conditional mutagenesis in zebrafish germ cells using Tol2 transposon and a CRISPR-Cas9-based plasmid system

STAR Protoc. 2024 Dec 20;6(1):103516. doi: 10.1016/j.xpro.2024.103516. Online ahead of print.

Authors

Yixuan Hu¹, Yutao Chen¹, Yu Zhang¹, Zhiquan Liu¹, Jianzhen Li²

Affiliations

¹ College of Life Sciences, Northwest Normal University, Lanzhou 730070, China.
² College of Life Sciences, Northwest Normal University, Lanzhou 730070, China. Electronic address: lijianzhen@nwnu.edu.cn.

PMID: 39709609
DOI: 10.1016/j.xpro.2024.103516

Abstract

Here, we present a protocol for conditional mutagenesis in zebrafish germ cells using Tol2 transposon and a CRISPR-Cas9-based plasmid system. We describe steps for conditional mutagenesis plasmid construction, zebrafish embryo microinjection, and screening for green fluorescence in the heart. This protocol is simple to execute, time efficient, and multifunctional, enabling the disruption of genes in zebrafish germ cells to be conducted with ease. For complete details on the use and execution of this protocol, please refer to Hu et al.¹.

Keywords: CRISPR; developmental biology; stem cells.