Precise and efficient genome editing in zebrafish using the CRISPR/Cas9 system
The introduction of engineered site-specific DNA endonucleases has brought precise genome editing in many model organisms and human cells into the realm of possibility. In zebrafish, loss-of-function alleles have been successfully produced; however, germ line transmission of functional targeted knoc...
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| Veröffentlicht in: | Development (Cambridge) 2014-12, Vol.141 (24), p.4827-4830 |
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| creator | Irion, Uwe Krauss, Jana Nüsslein-Volhard, Christiane |
| description | The introduction of engineered site-specific DNA endonucleases has brought precise genome editing in many model organisms and human cells into the realm of possibility. In zebrafish, loss-of-function alleles have been successfully produced; however, germ line transmission of functional targeted knock-ins of protein tags or of SNP exchanges have not been reported. Here we show by phenotypic rescue that the CRISPR/Cas system can be used to target and repair a premature stop codon at the albino (alb) locus in zebrafish with high efficiency and precision. Using circular donor DNA containing CRISPR target sites we obtain close to 50% of larvae with precise homology-directed repair of the alb(b4) mutation, a small fraction of which transmitted the repaired allele in the germ line to the next generation (3/28 adult fish). The in vivo demonstration of germ line transmission of a precise SNP exchange in zebrafish underscores its suitability as a model for genetic research. |
| doi_str_mv | 10.1242/dev.115584 |
| format | Article |
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In zebrafish, loss-of-function alleles have been successfully produced; however, germ line transmission of functional targeted knock-ins of protein tags or of SNP exchanges have not been reported. Here we show by phenotypic rescue that the CRISPR/Cas system can be used to target and repair a premature stop codon at the albino (alb) locus in zebrafish with high efficiency and precision. Using circular donor DNA containing CRISPR target sites we obtain close to 50% of larvae with precise homology-directed repair of the alb(b4) mutation, a small fraction of which transmitted the repaired allele in the germ line to the next generation (3/28 adult fish). The in vivo demonstration of germ line transmission of a precise SNP exchange in zebrafish underscores its suitability as a model for genetic research.</description><identifier>ISSN: 0950-1991</identifier><identifier>EISSN: 1477-9129</identifier><identifier>DOI: 10.1242/dev.115584</identifier><identifier>PMID: 25411213</identifier><language>eng</language><publisher>England: The Company of Biologists</publisher><subject>Animals ; Clustered Regularly Interspaced Short Palindromic Repeats - genetics ; Codon, Nonsense - genetics ; CRISPR-Associated Proteins - genetics ; Danio rerio ; DNA Primers - genetics ; DNA Repair - genetics ; DNA, Circular - genetics ; Genetic Engineering - methods ; Genome - genetics ; Genotype ; Membrane Transport Proteins - genetics ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide - genetics ; Techniques and Resources ; Zebrafish - genetics ; Zebrafish Proteins - genetics</subject><ispartof>Development (Cambridge), 2014-12, Vol.141 (24), p.4827-4830</ispartof><rights>2014. 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In zebrafish, loss-of-function alleles have been successfully produced; however, germ line transmission of functional targeted knock-ins of protein tags or of SNP exchanges have not been reported. Here we show by phenotypic rescue that the CRISPR/Cas system can be used to target and repair a premature stop codon at the albino (alb) locus in zebrafish with high efficiency and precision. Using circular donor DNA containing CRISPR target sites we obtain close to 50% of larvae with precise homology-directed repair of the alb(b4) mutation, a small fraction of which transmitted the repaired allele in the germ line to the next generation (3/28 adult fish). The in vivo demonstration of germ line transmission of a precise SNP exchange in zebrafish underscores its suitability as a model for genetic research.</description><subject>Animals</subject><subject>Clustered Regularly Interspaced Short Palindromic Repeats - genetics</subject><subject>Codon, Nonsense - genetics</subject><subject>CRISPR-Associated Proteins - genetics</subject><subject>Danio rerio</subject><subject>DNA Primers - genetics</subject><subject>DNA Repair - genetics</subject><subject>DNA, Circular - genetics</subject><subject>Genetic Engineering - methods</subject><subject>Genome - genetics</subject><subject>Genotype</subject><subject>Membrane Transport Proteins - genetics</subject><subject>Polymerase Chain Reaction</subject><subject>Polymorphism, Single Nucleotide - genetics</subject><subject>Techniques and Resources</subject><subject>Zebrafish - genetics</subject><subject>Zebrafish Proteins - genetics</subject><issn>0950-1991</issn><issn>1477-9129</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkUtLw0AUhQdRbK1u_AEySxHSziuZzEaQ4KNQtFRdD8nkph1pkppJCvXXO6W16MrVhXs-Dufeg9AlJUPKBBvlsB5SGoaxOEJ9KqQMFGXqGPWJCklAlaI9dObcByGER1Keoh4LBaWM8j56njZgrAOcVjmGorDGQtXiOVR1CRhy29pqjm2FvyBr0sK6Be7cdtUuACez8et0NkpSp7DbuBbKc3RSpEsHF_s5QO8P92_JUzB5eRwnd5PA-HhtUEQmpDEhuSo454wwnjEZES5jUCyOKeeGQ2QUEwDGxKSAjLOIg6A5FV7jA3S78111WQm58ZmbdKlXjS3TZqPr1Oq_SmUXel6vtWBKMSm8wfXeoKk_O3CtLq0zsFymFdSd0z5dLIlUEfsfjbynfz6XHr3ZoaapnWugOCSiRG-70r4rvevKw1e_bzigP-Xwb4SSjkI</recordid><startdate>20141215</startdate><enddate>20141215</enddate><creator>Irion, Uwe</creator><creator>Krauss, Jana</creator><creator>Nüsslein-Volhard, Christiane</creator><general>The Company of Biologists</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>5PM</scope></search><sort><creationdate>20141215</creationdate><title>Precise and efficient genome editing in zebrafish using the CRISPR/Cas9 system</title><author>Irion, Uwe ; Krauss, Jana ; Nüsslein-Volhard, Christiane</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c477t-f6c51800d9f3332023b2760378e9288133c3e6c924eecc80feb3263e41d143c33</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Animals</topic><topic>Clustered Regularly Interspaced Short Palindromic Repeats - genetics</topic><topic>Codon, Nonsense - genetics</topic><topic>CRISPR-Associated Proteins - genetics</topic><topic>Danio rerio</topic><topic>DNA Primers - genetics</topic><topic>DNA Repair - genetics</topic><topic>DNA, Circular - genetics</topic><topic>Genetic Engineering - methods</topic><topic>Genome - genetics</topic><topic>Genotype</topic><topic>Membrane Transport Proteins - genetics</topic><topic>Polymerase Chain Reaction</topic><topic>Polymorphism, Single Nucleotide - genetics</topic><topic>Techniques and Resources</topic><topic>Zebrafish - genetics</topic><topic>Zebrafish Proteins - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Irion, Uwe</creatorcontrib><creatorcontrib>Krauss, Jana</creatorcontrib><creatorcontrib>Nüsslein-Volhard, Christiane</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Development (Cambridge)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Irion, Uwe</au><au>Krauss, Jana</au><au>Nüsslein-Volhard, Christiane</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Precise and efficient genome editing in zebrafish using the CRISPR/Cas9 system</atitle><jtitle>Development (Cambridge)</jtitle><addtitle>Development</addtitle><date>2014-12-15</date><risdate>2014</risdate><volume>141</volume><issue>24</issue><spage>4827</spage><epage>4830</epage><pages>4827-4830</pages><issn>0950-1991</issn><eissn>1477-9129</eissn><abstract>The introduction of engineered site-specific DNA endonucleases has brought precise genome editing in many model organisms and human cells into the realm of possibility. In zebrafish, loss-of-function alleles have been successfully produced; however, germ line transmission of functional targeted knock-ins of protein tags or of SNP exchanges have not been reported. Here we show by phenotypic rescue that the CRISPR/Cas system can be used to target and repair a premature stop codon at the albino (alb) locus in zebrafish with high efficiency and precision. Using circular donor DNA containing CRISPR target sites we obtain close to 50% of larvae with precise homology-directed repair of the alb(b4) mutation, a small fraction of which transmitted the repaired allele in the germ line to the next generation (3/28 adult fish). The in vivo demonstration of germ line transmission of a precise SNP exchange in zebrafish underscores its suitability as a model for genetic research.</abstract><cop>England</cop><pub>The Company of Biologists</pub><pmid>25411213</pmid><doi>10.1242/dev.115584</doi><tpages>4</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0950-1991 |
| ispartof | Development (Cambridge), 2014-12, Vol.141 (24), p.4827-4830 |
| issn | 0950-1991 1477-9129 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_4299274 |
| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection; Company of Biologists |
| subjects | Animals Clustered Regularly Interspaced Short Palindromic Repeats - genetics Codon, Nonsense - genetics CRISPR-Associated Proteins - genetics Danio rerio DNA Primers - genetics DNA Repair - genetics DNA, Circular - genetics Genetic Engineering - methods Genome - genetics Genotype Membrane Transport Proteins - genetics Polymerase Chain Reaction Polymorphism, Single Nucleotide - genetics Techniques and Resources Zebrafish - genetics Zebrafish Proteins - genetics |
| title | Precise and efficient genome editing in zebrafish using the CRISPR/Cas9 system |
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