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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| Format: | Article |
| Language: | English |
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Elsevier
2025-03-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724006816 |
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| _version_ | 1846113101572734976 |
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| author | Yixuan Hu Yutao Chen Yu Zhang Zhiquan Liu Jianzhen Li |
| author_facet | Yixuan Hu Yutao Chen Yu Zhang Zhiquan Liu Jianzhen Li |
| author_sort | Yixuan Hu |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-dec932bb87b545808de28df7f2c974e9 |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-dec932bb87b545808de28df7f2c974e92024-12-22T05:29:47ZengElsevierSTAR Protocols2666-16672025-03-0161103516Protocol for conditional mutagenesis in zebrafish germ cells using Tol2 transposon and a CRISPR-Cas9-based plasmid systemYixuan Hu0Yutao Chen1Yu Zhang2Zhiquan Liu3Jianzhen Li4College of Life Sciences, Northwest Normal University, Lanzhou 730070, ChinaCollege of Life Sciences, Northwest Normal University, Lanzhou 730070, ChinaCollege of Life Sciences, Northwest Normal University, Lanzhou 730070, ChinaCollege of Life Sciences, Northwest Normal University, Lanzhou 730070, ChinaCollege of Life Sciences, Northwest Normal University, Lanzhou 730070, China; Corresponding authorSummary: 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.http://www.sciencedirect.com/science/article/pii/S2666166724006816developmental biologyCRISPRstem cells |
| spellingShingle | Yixuan Hu Yutao Chen Yu Zhang Zhiquan Liu Jianzhen Li Protocol for conditional mutagenesis in zebrafish germ cells using Tol2 transposon and a CRISPR-Cas9-based plasmid system STAR Protocols developmental biology CRISPR stem cells |
| title | Protocol for conditional mutagenesis in zebrafish germ cells using Tol2 transposon and a CRISPR-Cas9-based plasmid system |
| title_full | Protocol for conditional mutagenesis in zebrafish germ cells using Tol2 transposon and a CRISPR-Cas9-based plasmid system |
| title_fullStr | Protocol for conditional mutagenesis in zebrafish germ cells using Tol2 transposon and a CRISPR-Cas9-based plasmid system |
| title_full_unstemmed | Protocol for conditional mutagenesis in zebrafish germ cells using Tol2 transposon and a CRISPR-Cas9-based plasmid system |
| title_short | Protocol for conditional mutagenesis in zebrafish germ cells using Tol2 transposon and a CRISPR-Cas9-based plasmid system |
| title_sort | protocol for conditional mutagenesis in zebrafish germ cells using tol2 transposon and a crispr cas9 based plasmid system |
| topic | developmental biology CRISPR stem cells |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724006816 |
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