CRISPR/Cas9 System-Mediated Gene Editing in the Fujian Oysters (Crassostrea angulate) by Electroporation
The Fujian oyster (Crassostrea angulate) is an important marine bivalve mollusk with high economic value. Gene function research and gene editing techniques have broad application prospects in oyster. The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (...
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ftdoajarticles:oai:doaj.org/article:bee709975cc24123a45732e0cc303963 2023-05-15T17:54:19+02:00 CRISPR/Cas9 System-Mediated Gene Editing in the Fujian Oysters (Crassostrea angulate) by Electroporation Kaidi Jin Baolu Zhang Qianqian Jin Zhongqiang Cai Lei Wei Xiaomei Wang Yanxin Zheng Baoyu Huang Meiwei Zhang Yitao Qi Yaqiong Liu Xiaotong Wang 2021-10-01T00:00:00Z https://doi.org/10.3389/fmars.2021.763470 https://doaj.org/article/bee709975cc24123a45732e0cc303963 EN eng Frontiers Media S.A. https://www.frontiersin.org/articles/10.3389/fmars.2021.763470/full https://doaj.org/toc/2296-7745 2296-7745 doi:10.3389/fmars.2021.763470 https://doaj.org/article/bee709975cc24123a45732e0cc303963 Frontiers in Marine Science, Vol 8 (2021) CRISPR/Cas9 electroporation Crassostrea angulate gene editing mollusk Science Q General. Including nature conservation geographical distribution QH1-199.5 article 2021 ftdoajarticles https://doi.org/10.3389/fmars.2021.763470 2022-12-31T10:37:01Z The Fujian oyster (Crassostrea angulate) is an important marine bivalve mollusk with high economic value. Gene function research and gene editing techniques have broad application prospects in oyster. The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system has been widely used for genome engineering in many species. CRISPR-mediated gene editing has also been used successfully in the Pacific oyster through direct delivery of the CRISPR/Cas9 components into oyster embryos by microinjection. However, the low throughput and operational difficulties associated with microinjection is one of the factors limiting the widespread application of CRISPR/Cas9 in oysters. In this study, we attempted to deliver the CRISPR/Cas9-system into the embryos of C. angulate by electroporation. An all-in-one CRISPR/Cas9 vector plasmid was used as CRISPR/Cas9 system in this study. Electroporation was carried out using both eggs and blastula larvae. A large number of larvae became malformed or die after electroporation. A single base substitution mutation was detected in the D-larvae developed from electroporated eggs. Our results demonstrate that the CRISPR/Cas9 system can be delivered into embryos of C. angulate for gene editing by electroporation, which provides a reference and will further contribute to the future application of electroporation in mollusks. Article in Journal/Newspaper Pacific oyster Directory of Open Access Journals: DOAJ Articles Pacific Frontiers in Marine Science 8 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
CRISPR/Cas9 electroporation Crassostrea angulate gene editing mollusk Science Q General. Including nature conservation geographical distribution QH1-199.5 |
spellingShingle |
CRISPR/Cas9 electroporation Crassostrea angulate gene editing mollusk Science Q General. Including nature conservation geographical distribution QH1-199.5 Kaidi Jin Baolu Zhang Qianqian Jin Zhongqiang Cai Lei Wei Xiaomei Wang Yanxin Zheng Baoyu Huang Meiwei Zhang Yitao Qi Yaqiong Liu Xiaotong Wang CRISPR/Cas9 System-Mediated Gene Editing in the Fujian Oysters (Crassostrea angulate) by Electroporation |
topic_facet |
CRISPR/Cas9 electroporation Crassostrea angulate gene editing mollusk Science Q General. Including nature conservation geographical distribution QH1-199.5 |
description |
The Fujian oyster (Crassostrea angulate) is an important marine bivalve mollusk with high economic value. Gene function research and gene editing techniques have broad application prospects in oyster. The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system has been widely used for genome engineering in many species. CRISPR-mediated gene editing has also been used successfully in the Pacific oyster through direct delivery of the CRISPR/Cas9 components into oyster embryos by microinjection. However, the low throughput and operational difficulties associated with microinjection is one of the factors limiting the widespread application of CRISPR/Cas9 in oysters. In this study, we attempted to deliver the CRISPR/Cas9-system into the embryos of C. angulate by electroporation. An all-in-one CRISPR/Cas9 vector plasmid was used as CRISPR/Cas9 system in this study. Electroporation was carried out using both eggs and blastula larvae. A large number of larvae became malformed or die after electroporation. A single base substitution mutation was detected in the D-larvae developed from electroporated eggs. Our results demonstrate that the CRISPR/Cas9 system can be delivered into embryos of C. angulate for gene editing by electroporation, which provides a reference and will further contribute to the future application of electroporation in mollusks. |
format |
Article in Journal/Newspaper |
author |
Kaidi Jin Baolu Zhang Qianqian Jin Zhongqiang Cai Lei Wei Xiaomei Wang Yanxin Zheng Baoyu Huang Meiwei Zhang Yitao Qi Yaqiong Liu Xiaotong Wang |
author_facet |
Kaidi Jin Baolu Zhang Qianqian Jin Zhongqiang Cai Lei Wei Xiaomei Wang Yanxin Zheng Baoyu Huang Meiwei Zhang Yitao Qi Yaqiong Liu Xiaotong Wang |
author_sort |
Kaidi Jin |
title |
CRISPR/Cas9 System-Mediated Gene Editing in the Fujian Oysters (Crassostrea angulate) by Electroporation |
title_short |
CRISPR/Cas9 System-Mediated Gene Editing in the Fujian Oysters (Crassostrea angulate) by Electroporation |
title_full |
CRISPR/Cas9 System-Mediated Gene Editing in the Fujian Oysters (Crassostrea angulate) by Electroporation |
title_fullStr |
CRISPR/Cas9 System-Mediated Gene Editing in the Fujian Oysters (Crassostrea angulate) by Electroporation |
title_full_unstemmed |
CRISPR/Cas9 System-Mediated Gene Editing in the Fujian Oysters (Crassostrea angulate) by Electroporation |
title_sort |
crispr/cas9 system-mediated gene editing in the fujian oysters (crassostrea angulate) by electroporation |
publisher |
Frontiers Media S.A. |
publishDate |
2021 |
url |
https://doi.org/10.3389/fmars.2021.763470 https://doaj.org/article/bee709975cc24123a45732e0cc303963 |
geographic |
Pacific |
geographic_facet |
Pacific |
genre |
Pacific oyster |
genre_facet |
Pacific oyster |
op_source |
Frontiers in Marine Science, Vol 8 (2021) |
op_relation |
https://www.frontiersin.org/articles/10.3389/fmars.2021.763470/full https://doaj.org/toc/2296-7745 2296-7745 doi:10.3389/fmars.2021.763470 https://doaj.org/article/bee709975cc24123a45732e0cc303963 |
op_doi |
https://doi.org/10.3389/fmars.2021.763470 |
container_title |
Frontiers in Marine Science |
container_volume |
8 |
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1766162071883350016 |