Protocol for CRISPR-Cas9-mediated genome editing to study spermatogenesis in Caenorhabditis elegans
Summary: Gene silencing by P-element-induced wimpy testis-interacting RNAs is a mechanism to maintain genome integrity in germ cells. Here, we present a protocol for knockin or knockout editing of male germline genome mediated by CRISPR-Cas9 technology in Caenorhabditis elegans. We describe steps fo...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-12-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723006871 |
| _version_ | 1797627088918806528 |
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| author | Peng Wang Zheng Cao Qiushi Wang Xiaojing Ma Ning Wang Lianwan Chen Yanmei Zhao Long Miao |
| author_facet | Peng Wang Zheng Cao Qiushi Wang Xiaojing Ma Ning Wang Lianwan Chen Yanmei Zhao Long Miao |
| author_sort | Peng Wang |
| collection | DOAJ |
| description | Summary: Gene silencing by P-element-induced wimpy testis-interacting RNAs is a mechanism to maintain genome integrity in germ cells. Here, we present a protocol for knockin or knockout editing of male germline genome mediated by CRISPR-Cas9 technology in Caenorhabditis elegans. We describe steps for constructing edited plasmids, microinjecting worms with these plasmids, and screening edited worms. We then detail procedures for dissecting released sperm and their observation with fluorescence microscopy. Engineered worms provide a model for studying hermaphrodite/male fertility or protein localization in vivo.For complete details on the use and execution of this protocol, please refer to Wang et al. (2021).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-11T10:19:23Z |
| format | Article |
| id | doaj.art-2e4547a59a33408da1c9ce8cb0257fd7 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-11T10:19:23Z |
| publishDate | 2023-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-2e4547a59a33408da1c9ce8cb0257fd72023-11-16T06:10:57ZengElsevierSTAR Protocols2666-16672023-12-0144102720Protocol for CRISPR-Cas9-mediated genome editing to study spermatogenesis in Caenorhabditis elegansPeng Wang0Zheng Cao1Qiushi Wang2Xiaojing Ma3Ning Wang4Lianwan Chen5Yanmei Zhao6Long Miao7Key Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100059, ChinaKey Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, ChinaKey Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100059, ChinaKey Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100059, ChinaKey Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100059, ChinaKey Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, ChinaKey Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; Corresponding authorKey Laboratory of RNA Biology, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100059, China; Center for Biological Imaging, Core Facilities for Protein Science, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; MOE Key Laboratory of Cell Proliferation and Regulation Biology, College of Life Sciences, Beijing Normal University, Beijing 100875, China; Corresponding authorSummary: Gene silencing by P-element-induced wimpy testis-interacting RNAs is a mechanism to maintain genome integrity in germ cells. Here, we present a protocol for knockin or knockout editing of male germline genome mediated by CRISPR-Cas9 technology in Caenorhabditis elegans. We describe steps for constructing edited plasmids, microinjecting worms with these plasmids, and screening edited worms. We then detail procedures for dissecting released sperm and their observation with fluorescence microscopy. Engineered worms provide a model for studying hermaphrodite/male fertility or protein localization in vivo.For complete details on the use and execution of this protocol, please refer to Wang et al. (2021).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/S2666166723006871Cell BiologyDevelopmental biologyGeneticsCRISPR |
| spellingShingle | Peng Wang Zheng Cao Qiushi Wang Xiaojing Ma Ning Wang Lianwan Chen Yanmei Zhao Long Miao Protocol for CRISPR-Cas9-mediated genome editing to study spermatogenesis in Caenorhabditis elegans STAR Protocols Cell Biology Developmental biology Genetics CRISPR |
| title | Protocol for CRISPR-Cas9-mediated genome editing to study spermatogenesis in Caenorhabditis elegans |
| title_full | Protocol for CRISPR-Cas9-mediated genome editing to study spermatogenesis in Caenorhabditis elegans |
| title_fullStr | Protocol for CRISPR-Cas9-mediated genome editing to study spermatogenesis in Caenorhabditis elegans |
| title_full_unstemmed | Protocol for CRISPR-Cas9-mediated genome editing to study spermatogenesis in Caenorhabditis elegans |
| title_short | Protocol for CRISPR-Cas9-mediated genome editing to study spermatogenesis in Caenorhabditis elegans |
| title_sort | protocol for crispr cas9 mediated genome editing to study spermatogenesis in caenorhabditis elegans |
| topic | Cell Biology Developmental biology Genetics CRISPR |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723006871 |
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