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

Summary: 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 hear...

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Bibliographic Details
Main Authors: Yixuan Hu, Yutao Chen, Yu Zhang, Zhiquan Liu, Jianzhen Li
Format: Article
Language:English
Published: Elsevier 2025-03-01
Series:STAR Protocols
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666166724006816
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Summary:Summary: 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 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
ISSN:2666-1667