A detailed procedure for CRISPR/Cas9-mediated gene editing in tilapia
Reverse genetics approaches are critical for uncovering complex biological processes and genetic engineering. Clustered regularly interspaced short palindromic repeats (CRISPR)–CRISPR-associated (Cas) system (CRISPR/Cas9) has been widely used for generating mutants in a large number of species. The...
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| Veröffentlicht in: | Hydrobiologia 2021-09, Vol.848 (16), p.3865-3881 |
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| creator | Li, Minghui Dai, Shengfei Liu, Xingyong Xiao, Hesheng Wang, Deshou |
| description | Reverse genetics approaches are critical for uncovering complex biological processes and genetic engineering. Clustered regularly interspaced short palindromic repeats (CRISPR)–CRISPR-associated (Cas) system (CRISPR/Cas9) has been widely used for generating mutants in a large number of species. The Nile tilapia (
Oreochromis niloticus
), a gonochoristic teleost with XX/XY sex determination system, is a good model for studying fish sex determination and reproduction. Here, we describe a detailed procedure for gene mutation in tilapia by CRISPR/Cas9. This protocol covers selection of target sites, in vitro RNA transcription, artificial insemination and microinjection of one cell stage embryos. We also provide details for detection of somatic and germline transmitted mutations, fast establishment of mutant lines and discuss some practical advices. This protocol will facilitate broader applications of CRISPR/Cas9 in studies of tilapia as well as other aquaculture fishes. |
| doi_str_mv | 10.1007/s10750-020-04414-8 |
| format | Article |
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Oreochromis niloticus
), a gonochoristic teleost with XX/XY sex determination system, is a good model for studying fish sex determination and reproduction. Here, we describe a detailed procedure for gene mutation in tilapia by CRISPR/Cas9. This protocol covers selection of target sites, in vitro RNA transcription, artificial insemination and microinjection of one cell stage embryos. We also provide details for detection of somatic and germline transmitted mutations, fast establishment of mutant lines and discuss some practical advices. This protocol will facilitate broader applications of CRISPR/Cas9 in studies of tilapia as well as other aquaculture fishes.</description><identifier>ISSN: 0018-8158</identifier><identifier>EISSN: 1573-5117</identifier><identifier>DOI: 10.1007/s10750-020-04414-8</identifier><language>eng</language><publisher>Cham: Springer International Publishing</publisher><subject>Advances in Cichlid Research IV ; Analysis ; Animal embryos ; Aquaculture ; Artificial insemination ; Biological activity ; Biological fertilization ; Biomedical and Life Sciences ; CRISPR ; Ecology ; Embryos ; Fish ; Fish reproduction ; Fishes ; Freshwater & Marine Ecology ; Freshwater fishes ; Gene editing ; Gene mutations ; Genetic aspects ; Genetic engineering ; Genetic modification ; Genetic research ; Genetic transcription ; Genetically modified organisms ; Genetics ; Life Sciences ; Marine fishes ; Microinjection ; Mutants ; Mutation ; Oreochromis niloticus ; Point mutation ; Ponds ; Procedures ; Reproduction (biology) ; RNA ; Sex ; Sex determination ; Tilapia ; Transcription ; Zoology</subject><ispartof>Hydrobiologia, 2021-09, Vol.848 (16), p.3865-3881</ispartof><rights>Springer Nature Switzerland AG 2020</rights><rights>COPYRIGHT 2021 Springer</rights><rights>Springer Nature Switzerland AG 2020.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c392t-13517970b69cffce0f5791529cdb33fba7c0ef18bee3c151ba9d52e4a8109b4b3</citedby><cites>FETCH-LOGICAL-c392t-13517970b69cffce0f5791529cdb33fba7c0ef18bee3c151ba9d52e4a8109b4b3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s10750-020-04414-8$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s10750-020-04414-8$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27903,27904,41467,42536,51297</link.rule.ids></links><search><creatorcontrib>Li, Minghui</creatorcontrib><creatorcontrib>Dai, Shengfei</creatorcontrib><creatorcontrib>Liu, Xingyong</creatorcontrib><creatorcontrib>Xiao, Hesheng</creatorcontrib><creatorcontrib>Wang, Deshou</creatorcontrib><title>A detailed procedure for CRISPR/Cas9-mediated gene editing in tilapia</title><title>Hydrobiologia</title><addtitle>Hydrobiologia</addtitle><description>Reverse genetics approaches are critical for uncovering complex biological processes and genetic engineering. Clustered regularly interspaced short palindromic repeats (CRISPR)–CRISPR-associated (Cas) system (CRISPR/Cas9) has been widely used for generating mutants in a large number of species. The Nile tilapia (
Oreochromis niloticus
), a gonochoristic teleost with XX/XY sex determination system, is a good model for studying fish sex determination and reproduction. Here, we describe a detailed procedure for gene mutation in tilapia by CRISPR/Cas9. This protocol covers selection of target sites, in vitro RNA transcription, artificial insemination and microinjection of one cell stage embryos. We also provide details for detection of somatic and germline transmitted mutations, fast establishment of mutant lines and discuss some practical advices. This protocol will facilitate broader applications of CRISPR/Cas9 in studies of tilapia as well as other aquaculture fishes.</description><subject>Advances in Cichlid Research IV</subject><subject>Analysis</subject><subject>Animal embryos</subject><subject>Aquaculture</subject><subject>Artificial insemination</subject><subject>Biological activity</subject><subject>Biological fertilization</subject><subject>Biomedical and Life Sciences</subject><subject>CRISPR</subject><subject>Ecology</subject><subject>Embryos</subject><subject>Fish</subject><subject>Fish reproduction</subject><subject>Fishes</subject><subject>Freshwater & Marine Ecology</subject><subject>Freshwater fishes</subject><subject>Gene editing</subject><subject>Gene mutations</subject><subject>Genetic aspects</subject><subject>Genetic engineering</subject><subject>Genetic modification</subject><subject>Genetic research</subject><subject>Genetic transcription</subject><subject>Genetically modified organisms</subject><subject>Genetics</subject><subject>Life Sciences</subject><subject>Marine fishes</subject><subject>Microinjection</subject><subject>Mutants</subject><subject>Mutation</subject><subject>Oreochromis niloticus</subject><subject>Point mutation</subject><subject>Ponds</subject><subject>Procedures</subject><subject>Reproduction (biology)</subject><subject>RNA</subject><subject>Sex</subject><subject>Sex 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tilapia</atitle><jtitle>Hydrobiologia</jtitle><stitle>Hydrobiologia</stitle><date>2021-09-01</date><risdate>2021</risdate><volume>848</volume><issue>16</issue><spage>3865</spage><epage>3881</epage><pages>3865-3881</pages><issn>0018-8158</issn><eissn>1573-5117</eissn><abstract>Reverse genetics approaches are critical for uncovering complex biological processes and genetic engineering. Clustered regularly interspaced short palindromic repeats (CRISPR)–CRISPR-associated (Cas) system (CRISPR/Cas9) has been widely used for generating mutants in a large number of species. The Nile tilapia (
Oreochromis niloticus
), a gonochoristic teleost with XX/XY sex determination system, is a good model for studying fish sex determination and reproduction. Here, we describe a detailed procedure for gene mutation in tilapia by CRISPR/Cas9. This protocol covers selection of target sites, in vitro RNA transcription, artificial insemination and microinjection of one cell stage embryos. We also provide details for detection of somatic and germline transmitted mutations, fast establishment of mutant lines and discuss some practical advices. This protocol will facilitate broader applications of CRISPR/Cas9 in studies of tilapia as well as other aquaculture fishes.</abstract><cop>Cham</cop><pub>Springer International Publishing</pub><doi>10.1007/s10750-020-04414-8</doi><tpages>17</tpages></addata></record> |
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| subjects | Advances in Cichlid Research IV Analysis Animal embryos Aquaculture Artificial insemination Biological activity Biological fertilization Biomedical and Life Sciences CRISPR Ecology Embryos Fish Fish reproduction Fishes Freshwater & Marine Ecology Freshwater fishes Gene editing Gene mutations Genetic aspects Genetic engineering Genetic modification Genetic research Genetic transcription Genetically modified organisms Genetics Life Sciences Marine fishes Microinjection Mutants Mutation Oreochromis niloticus Point mutation Ponds Procedures Reproduction (biology) RNA Sex Sex determination Tilapia Transcription Zoology |
| title | A detailed procedure for CRISPR/Cas9-mediated gene editing in tilapia |
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