Correction of the auditory phenotype in C57BL/6N mice via CRISPR/Cas9-mediated homology directed repair View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2016-02-15

AUTHORS

Joffrey Mianné, Lauren Chessum, Saumya Kumar, Carlos Aguilar, Gemma Codner, Marie Hutchison, Andrew Parker, Ann-Marie Mallon, Sara Wells, Michelle M. Simon, Lydia Teboul, Steve D. M. Brown, Michael R. Bowl

ABSTRACT

BackgroundNuclease-based technologies have been developed that enable targeting of specific DNA sequences directly in the zygote. These approaches provide an opportunity to modify the genomes of inbred mice, and allow the removal of strain-specific mutations that confound phenotypic assessment. One such mutation is the Cdh23ahl allele, present in several commonly used inbred mouse strains, which predisposes to age-related progressive hearing loss.ResultsWe have used targeted CRISPR/Cas9-mediated homology directed repair (HDR) to correct the Cdh23ahl allele directly in C57BL/6NTac zygotes. Employing offset-nicking Cas9 (D10A) nickase with paired RNA guides and a single-stranded oligonucleotide donor template we show that allele repair was successfully achieved. To investigate potential Cas9-mediated ‘off-target’ mutations in our corrected mouse, we undertook whole-genome sequencing and assessed the ‘off-target’ sites predicted for the guide RNAs (≤4 nucleotide mis-matches). No induced sequence changes were identified at any of these sites.Correction of the progressive hearing loss phenotype was demonstrated using auditory-evoked brainstem response testing of mice at 24 and 36 weeks of age, and rescue of the progressive loss of sensory hair cell stereocilia bundles was confirmed using scanning electron microscopy of dissected cochleae from 36-week-old mice.ConclusionsCRISPR/Cas9-mediated HDR has been successfully utilised to efficiently correct the Cdh23ahl allele in C57BL/6NTac mice, and rescue the associated auditory phenotype. The corrected mice described in this report will allow age-related auditory phenotyping studies to be undertaken using C57BL/6NTac-derived models, such as those generated by the International Mouse Phenotyping Consortium (IMPC) programme. More... »

PAGES

16

References to SciGraph publications

  • 2001-06-01. Spiral Ligament Pathology: A Major Aspect of Age-Related Cochlear Degeneration in C57BL/6 Mice in JOURNAL OF THE ASSOCIATION FOR RESEARCH IN OTOLARYNGOLOGY
  • 2010-01-07. A hearing and vestibular phenotyping pipeline to identify mouse mutants with hearing impairment in NATURE PROTOCOLS
  • 2011-09-14. Mouse genomic variation and its effect on phenotypes and gene regulation in NATURE
  • 2013-10-24. Genome engineering using the CRISPR-Cas9 system in NATURE PROTOCOLS
  • 2013-06-23. High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells in NATURE BIOTECHNOLOGY
  • 2012-09-01. The International Mouse Phenotyping Consortium: past and future perspectives on mouse phenotyping in MAMMALIAN GENOME
  • 2014-06-23. Rapid generation of mouse models with defined point mutations by the CRISPR/Cas9 system in SCIENTIFIC REPORTS
  • 2008-02-19. Quiet as a mouse: dissecting the molecular and genetic basis of hearing in NATURE REVIEWS GENETICS
  • 2009-05-15. Overexpression Transgenesis in Mouse: Pronuclear Injection in TRANSGENESIS TECHNIQUES
  • 2013-07-21. DNA targeting specificity of RNA-guided Cas9 nucleases in NATURE BIOTECHNOLOGY
  • 2014-03-02. Efficient genome modification by CRISPR-Cas9 nickase with minimal off-target effects in NATURE METHODS
  • 2014-06-18. Genome-wide association analysis demonstrates the highly polygenic character of age-related hearing impairment in EUROPEAN JOURNAL OF HUMAN GENETICS
  • 2010-01-13. A genome-wide association study for age-related hearing impairment in the Saami in EUROPEAN JOURNAL OF HUMAN GENETICS
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1186/s13073-016-0273-4

    DOI

    http://dx.doi.org/10.1186/s13073-016-0273-4

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1045564567

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/26876963


    Indexing Status Check whether this publication has been indexed by Scopus and Web Of Science using the SN Indexing Status Tool
    Incoming Citations Browse incoming citations for this publication using opencitations.net

    JSON-LD is the canonical representation for SciGraph data.

    TIP: You can open this SciGraph record using an external JSON-LD service: JSON-LD Playground Google SDTT

    [
      {
        "@context": "https://springernature.github.io/scigraph/jsonld/sgcontext.json", 
        "about": [
          {
            "id": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/06", 
            "inDefinedTermSet": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/", 
            "name": "Biological Sciences", 
            "type": "DefinedTerm"
          }, 
          {
            "id": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/0604", 
            "inDefinedTermSet": "http://purl.org/au-research/vocabulary/anzsrc-for/2008/", 
            "name": "Genetics", 
            "type": "DefinedTerm"
          }, 
          {
            "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
            "name": "Animals", 
            "type": "DefinedTerm"
          }, 
          {
            "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
            "name": "CRISPR-Cas Systems", 
            "type": "DefinedTerm"
          }, 
          {
            "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
            "name": "Cadherins", 
            "type": "DefinedTerm"
          }, 
          {
            "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
            "name": "Evoked Potentials, Auditory", 
            "type": "DefinedTerm"
          }, 
          {
            "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
            "name": "Genetic Therapy", 
            "type": "DefinedTerm"
          }, 
          {
            "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
            "name": "Hearing Loss", 
            "type": "DefinedTerm"
          }, 
          {
            "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
            "name": "Mice", 
            "type": "DefinedTerm"
          }, 
          {
            "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
            "name": "Mice, Inbred C57BL", 
            "type": "DefinedTerm"
          }, 
          {
            "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
            "name": "RNA, Guide", 
            "type": "DefinedTerm"
          }, 
          {
            "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
            "name": "Recombinational DNA Repair", 
            "type": "DefinedTerm"
          }, 
          {
            "inDefinedTermSet": "https://www.nlm.nih.gov/mesh/", 
            "name": "Stereocilia", 
            "type": "DefinedTerm"
          }
        ], 
        "author": [
          {
            "affiliation": {
              "alternateName": "Mary Lyon Centre, MRC Harwell, Harwell, OX11 0RD, Oxford, UK", 
              "id": "http://www.grid.ac/institutes/grid.420006.0", 
              "name": [
                "Mary Lyon Centre, MRC Harwell, Harwell, OX11 0RD, Oxford, UK"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Miann\u00e9", 
            "givenName": "Joffrey", 
            "id": "sg:person.0601633627.85", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0601633627.85"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "Mammalian Genetics Unit, MRC Harwell, Harwell, OX11 0RD, Oxford, UK", 
              "id": "http://www.grid.ac/institutes/grid.420006.0", 
              "name": [
                "Mammalian Genetics Unit, MRC Harwell, Harwell, OX11 0RD, Oxford, UK"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Chessum", 
            "givenName": "Lauren", 
            "id": "sg:person.0632760571.83", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0632760571.83"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "Mammalian Genetics Unit, MRC Harwell, Harwell, OX11 0RD, Oxford, UK", 
              "id": "http://www.grid.ac/institutes/grid.420006.0", 
              "name": [
                "Mammalian Genetics Unit, MRC Harwell, Harwell, OX11 0RD, Oxford, UK"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Kumar", 
            "givenName": "Saumya", 
            "id": "sg:person.01356107722.37", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01356107722.37"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "Mammalian Genetics Unit, MRC Harwell, Harwell, OX11 0RD, Oxford, UK", 
              "id": "http://www.grid.ac/institutes/grid.420006.0", 
              "name": [
                "Mammalian Genetics Unit, MRC Harwell, Harwell, OX11 0RD, Oxford, UK"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Aguilar", 
            "givenName": "Carlos", 
            "id": "sg:person.01317506032.68", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01317506032.68"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "Mary Lyon Centre, MRC Harwell, Harwell, OX11 0RD, Oxford, UK", 
              "id": "http://www.grid.ac/institutes/grid.420006.0", 
              "name": [
                "Mary Lyon Centre, MRC Harwell, Harwell, OX11 0RD, Oxford, UK"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Codner", 
            "givenName": "Gemma", 
            "id": "sg:person.0631215675.09", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0631215675.09"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "Mary Lyon Centre, MRC Harwell, Harwell, OX11 0RD, Oxford, UK", 
              "id": "http://www.grid.ac/institutes/grid.420006.0", 
              "name": [
                "Mary Lyon Centre, MRC Harwell, Harwell, OX11 0RD, Oxford, UK"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Hutchison", 
            "givenName": "Marie", 
            "id": "sg:person.01100424027.43", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01100424027.43"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "Mammalian Genetics Unit, MRC Harwell, Harwell, OX11 0RD, Oxford, UK", 
              "id": "http://www.grid.ac/institutes/grid.420006.0", 
              "name": [
                "Mammalian Genetics Unit, MRC Harwell, Harwell, OX11 0RD, Oxford, UK"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Parker", 
            "givenName": "Andrew", 
            "id": "sg:person.016020736737.08", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.016020736737.08"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "Mammalian Genetics Unit, MRC Harwell, Harwell, OX11 0RD, Oxford, UK", 
              "id": "http://www.grid.ac/institutes/grid.420006.0", 
              "name": [
                "Mammalian Genetics Unit, MRC Harwell, Harwell, OX11 0RD, Oxford, UK"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Mallon", 
            "givenName": "Ann-Marie", 
            "id": "sg:person.01337530626.28", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01337530626.28"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "Mary Lyon Centre, MRC Harwell, Harwell, OX11 0RD, Oxford, UK", 
              "id": "http://www.grid.ac/institutes/grid.420006.0", 
              "name": [
                "Mary Lyon Centre, MRC Harwell, Harwell, OX11 0RD, Oxford, UK"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Wells", 
            "givenName": "Sara", 
            "id": "sg:person.01244331330.82", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01244331330.82"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "Mammalian Genetics Unit, MRC Harwell, Harwell, OX11 0RD, Oxford, UK", 
              "id": "http://www.grid.ac/institutes/grid.420006.0", 
              "name": [
                "Mammalian Genetics Unit, MRC Harwell, Harwell, OX11 0RD, Oxford, UK"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Simon", 
            "givenName": "Michelle M.", 
            "id": "sg:person.01352202371.19", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01352202371.19"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "Mary Lyon Centre, MRC Harwell, Harwell, OX11 0RD, Oxford, UK", 
              "id": "http://www.grid.ac/institutes/grid.420006.0", 
              "name": [
                "Mary Lyon Centre, MRC Harwell, Harwell, OX11 0RD, Oxford, UK"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Teboul", 
            "givenName": "Lydia", 
            "id": "sg:person.01032576572.09", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01032576572.09"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "Mammalian Genetics Unit, MRC Harwell, Harwell, OX11 0RD, Oxford, UK", 
              "id": "http://www.grid.ac/institutes/grid.420006.0", 
              "name": [
                "Mammalian Genetics Unit, MRC Harwell, Harwell, OX11 0RD, Oxford, UK"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Brown", 
            "givenName": "Steve D. M.", 
            "id": "sg:person.012064305702.93", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.012064305702.93"
            ], 
            "type": "Person"
          }, 
          {
            "affiliation": {
              "alternateName": "Mammalian Genetics Unit, MRC Harwell, Harwell, OX11 0RD, Oxford, UK", 
              "id": "http://www.grid.ac/institutes/grid.420006.0", 
              "name": [
                "Mammalian Genetics Unit, MRC Harwell, Harwell, OX11 0RD, Oxford, UK"
              ], 
              "type": "Organization"
            }, 
            "familyName": "Bowl", 
            "givenName": "Michael R.", 
            "id": "sg:person.01047440473.56", 
            "sameAs": [
              "https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01047440473.56"
            ], 
            "type": "Person"
          }
        ], 
        "citation": [
          {
            "id": "sg:pub.10.1038/ejhg.2009.234", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1036164743", 
              "https://doi.org/10.1038/ejhg.2009.234"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1038/nbt.2623", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1009511624", 
              "https://doi.org/10.1038/nbt.2623"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1038/ejhg.2014.56", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1031381434", 
              "https://doi.org/10.1038/ejhg.2014.56"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1038/nmeth.2857", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1035757498", 
              "https://doi.org/10.1038/nmeth.2857"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1007/978-1-60327-019-9_8", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1016698235", 
              "https://doi.org/10.1007/978-1-60327-019-9_8"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1007/s101620010075", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1038584115", 
              "https://doi.org/10.1007/s101620010075"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1038/nbt.2647", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1012543463", 
              "https://doi.org/10.1038/nbt.2647"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1038/nature10413", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1041325874", 
              "https://doi.org/10.1038/nature10413"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1038/nrg2309", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1001777660", 
              "https://doi.org/10.1038/nrg2309"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1038/srep05396", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1011163022", 
              "https://doi.org/10.1038/srep05396"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1007/s00335-012-9427-x", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1009915802", 
              "https://doi.org/10.1007/s00335-012-9427-x"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1038/nprot.2013.143", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1016534425", 
              "https://doi.org/10.1038/nprot.2013.143"
            ], 
            "type": "CreativeWork"
          }, 
          {
            "id": "sg:pub.10.1038/nprot.2009.204", 
            "sameAs": [
              "https://app.dimensions.ai/details/publication/pub.1039038752", 
              "https://doi.org/10.1038/nprot.2009.204"
            ], 
            "type": "CreativeWork"
          }
        ], 
        "datePublished": "2016-02-15", 
        "datePublishedReg": "2016-02-15", 
        "description": "BackgroundNuclease-based technologies have been developed that enable targeting of specific DNA sequences directly in the zygote. These approaches provide an opportunity to modify the genomes of inbred mice, and allow the removal of strain-specific mutations that confound phenotypic assessment. One such mutation is the Cdh23ahl allele, present in several commonly used inbred mouse strains, which predisposes to age-related progressive hearing loss.ResultsWe have used targeted CRISPR/Cas9-mediated homology directed repair (HDR) to correct the Cdh23ahl allele directly in C57BL/6NTac zygotes. Employing offset-nicking Cas9 (D10A) nickase with paired RNA guides and a single-stranded oligonucleotide donor template we show that allele repair was successfully achieved. To investigate potential Cas9-mediated \u2018off-target\u2019 mutations in our corrected mouse, we undertook whole-genome sequencing and assessed the \u2018off-target\u2019 sites predicted for the guide RNAs (\u22644 nucleotide mis-matches). No induced sequence changes were identified at any of these sites.Correction of the progressive hearing loss phenotype was demonstrated using auditory-evoked brainstem response testing of mice at 24 and 36\u00a0weeks of age, and rescue of the progressive loss of sensory hair cell stereocilia bundles was confirmed using scanning electron microscopy of dissected cochleae from 36-week-old mice.ConclusionsCRISPR/Cas9-mediated HDR has been successfully utilised to efficiently correct the Cdh23ahl allele in C57BL/6NTac mice, and rescue the associated auditory phenotype. The corrected mice described in this report will allow age-related auditory phenotyping studies to be undertaken using C57BL/6NTac-derived models, such as those generated by the International Mouse Phenotyping Consortium (IMPC) programme.", 
        "genre": "article", 
        "id": "sg:pub.10.1186/s13073-016-0273-4", 
        "isAccessibleForFree": true, 
        "isFundedItemOf": [
          {
            "id": "sg:grant.7440373", 
            "type": "MonetaryGrant"
          }, 
          {
            "id": "sg:grant.2756236", 
            "type": "MonetaryGrant"
          }, 
          {
            "id": "sg:grant.5498354", 
            "type": "MonetaryGrant"
          }, 
          {
            "id": "sg:grant.2752996", 
            "type": "MonetaryGrant"
          }, 
          {
            "id": "sg:grant.2782647", 
            "type": "MonetaryGrant"
          }, 
          {
            "id": "sg:grant.7921244", 
            "type": "MonetaryGrant"
          }
        ], 
        "isPartOf": [
          {
            "id": "sg:journal.1040124", 
            "issn": [
              "1756-994X"
            ], 
            "name": "Genome Medicine", 
            "publisher": "Springer Nature", 
            "type": "Periodical"
          }, 
          {
            "issueNumber": "1", 
            "type": "PublicationIssue"
          }, 
          {
            "type": "PublicationVolume", 
            "volumeNumber": "8"
          }
        ], 
        "keywords": [
          "auditory phenotype", 
          "progressive hearing loss", 
          "weeks of age", 
          "hair cell stereocilia bundles", 
          "hearing loss phenotype", 
          "C57BL/6N mice", 
          "hearing loss", 
          "mice", 
          "progressive loss", 
          "mouse strains", 
          "response testing", 
          "stereocilia bundles", 
          "loss phenotype", 
          "whole-genome sequencing", 
          "strain-specific mutations", 
          "repair", 
          "auditory", 
          "phenotype", 
          "such mutations", 
          "phenotypic assessment", 
          "mutations", 
          "alleles", 
          "weeks", 
          "ResultsWe", 
          "CRISPR/", 
          "cochlea", 
          "C57BL/6NTac", 
          "age", 
          "target", 
          "sequence changes", 
          "report", 
          "loss", 
          "targeting", 
          "Consortium Program", 
          "zygotes", 
          "assessment", 
          "RNA", 
          "rescue", 
          "sequencing", 
          "testing", 
          "sites", 
          "study", 
          "correction", 
          "strains", 
          "changes", 
          "HDR", 
          "program", 
          "homology", 
          "bundles", 
          "removal", 
          "guide", 
          "specific DNA sequences", 
          "donor template", 
          "electron microscopy", 
          "opportunities", 
          "DNA sequences", 
          "Cas9 nickase", 
          "model", 
          "genome", 
          "guide RNA", 
          "microscopy", 
          "approach", 
          "RNA guides", 
          "sequence", 
          "nickase", 
          "template", 
          "technology"
        ], 
        "name": "Correction of the auditory phenotype in C57BL/6N mice via CRISPR/Cas9-mediated homology directed repair", 
        "pagination": "16", 
        "productId": [
          {
            "name": "dimensions_id", 
            "type": "PropertyValue", 
            "value": [
              "pub.1045564567"
            ]
          }, 
          {
            "name": "doi", 
            "type": "PropertyValue", 
            "value": [
              "10.1186/s13073-016-0273-4"
            ]
          }, 
          {
            "name": "pubmed_id", 
            "type": "PropertyValue", 
            "value": [
              "26876963"
            ]
          }
        ], 
        "sameAs": [
          "https://doi.org/10.1186/s13073-016-0273-4", 
          "https://app.dimensions.ai/details/publication/pub.1045564567"
        ], 
        "sdDataset": "articles", 
        "sdDatePublished": "2022-08-04T17:05", 
        "sdLicense": "https://scigraph.springernature.com/explorer/license/", 
        "sdPublisher": {
          "name": "Springer Nature - SN SciGraph project", 
          "type": "Organization"
        }, 
        "sdSource": "s3://com-springernature-scigraph/baseset/20220804/entities/gbq_results/article/article_713.jsonl", 
        "type": "ScholarlyArticle", 
        "url": "https://doi.org/10.1186/s13073-016-0273-4"
      }
    ]
     

    Download the RDF metadata as:  json-ld nt turtle xml License info

    HOW TO GET THIS DATA PROGRAMMATICALLY:

    JSON-LD is a popular format for linked data which is fully compatible with JSON.

    curl -H 'Accept: application/ld+json' 'https://scigraph.springernature.com/pub.10.1186/s13073-016-0273-4'

    N-Triples is a line-based linked data format ideal for batch operations.

    curl -H 'Accept: application/n-triples' 'https://scigraph.springernature.com/pub.10.1186/s13073-016-0273-4'

    Turtle is a human-readable linked data format.

    curl -H 'Accept: text/turtle' 'https://scigraph.springernature.com/pub.10.1186/s13073-016-0273-4'

    RDF/XML is a standard XML format for linked data.

    curl -H 'Accept: application/rdf+xml' 'https://scigraph.springernature.com/pub.10.1186/s13073-016-0273-4'


     

    This table displays all metadata directly associated to this object as RDF triples.

    321 TRIPLES      21 PREDICATES      116 URIs      95 LITERALS      18 BLANK NODES

    Subject Predicate Object
    1 sg:pub.10.1186/s13073-016-0273-4 schema:about N168bb13c4cae43c287817007334fa5af
    2 N4455285f41b84de9babd631ab1c42bad
    3 N5d045dc9a01c4343acf11b91dc46790a
    4 N611c89c6755e4952ba3c1cdc39b744fd
    5 N6d9bda0362d34954a3fea6f3cb031428
    6 Na74bfd5c84ca4aa58fbe7a9308b7dbbf
    7 Nd2268ab1f7fe45639da25153c423fc82
    8 Nd83e1602b624437ba6ac0be43016b347
    9 Ne88b89c32c9a4376984347d11a3dede0
    10 Nf648736c5d5446a9a4d114fb8805bd24
    11 Nf9d1c70f4cc74391bf674e313b5b7a22
    12 anzsrc-for:06
    13 anzsrc-for:0604
    14 schema:author N4ff3f3b99b164bddb8397643dc6d2971
    15 schema:citation sg:pub.10.1007/978-1-60327-019-9_8
    16 sg:pub.10.1007/s00335-012-9427-x
    17 sg:pub.10.1007/s101620010075
    18 sg:pub.10.1038/ejhg.2009.234
    19 sg:pub.10.1038/ejhg.2014.56
    20 sg:pub.10.1038/nature10413
    21 sg:pub.10.1038/nbt.2623
    22 sg:pub.10.1038/nbt.2647
    23 sg:pub.10.1038/nmeth.2857
    24 sg:pub.10.1038/nprot.2009.204
    25 sg:pub.10.1038/nprot.2013.143
    26 sg:pub.10.1038/nrg2309
    27 sg:pub.10.1038/srep05396
    28 schema:datePublished 2016-02-15
    29 schema:datePublishedReg 2016-02-15
    30 schema:description BackgroundNuclease-based technologies have been developed that enable targeting of specific DNA sequences directly in the zygote. These approaches provide an opportunity to modify the genomes of inbred mice, and allow the removal of strain-specific mutations that confound phenotypic assessment. One such mutation is the Cdh23ahl allele, present in several commonly used inbred mouse strains, which predisposes to age-related progressive hearing loss.ResultsWe have used targeted CRISPR/Cas9-mediated homology directed repair (HDR) to correct the Cdh23ahl allele directly in C57BL/6NTac zygotes. Employing offset-nicking Cas9 (D10A) nickase with paired RNA guides and a single-stranded oligonucleotide donor template we show that allele repair was successfully achieved. To investigate potential Cas9-mediated ‘off-target’ mutations in our corrected mouse, we undertook whole-genome sequencing and assessed the ‘off-target’ sites predicted for the guide RNAs (≤4 nucleotide mis-matches). No induced sequence changes were identified at any of these sites.Correction of the progressive hearing loss phenotype was demonstrated using auditory-evoked brainstem response testing of mice at 24 and 36 weeks of age, and rescue of the progressive loss of sensory hair cell stereocilia bundles was confirmed using scanning electron microscopy of dissected cochleae from 36-week-old mice.ConclusionsCRISPR/Cas9-mediated HDR has been successfully utilised to efficiently correct the Cdh23ahl allele in C57BL/6NTac mice, and rescue the associated auditory phenotype. The corrected mice described in this report will allow age-related auditory phenotyping studies to be undertaken using C57BL/6NTac-derived models, such as those generated by the International Mouse Phenotyping Consortium (IMPC) programme.
    31 schema:genre article
    32 schema:isAccessibleForFree true
    33 schema:isPartOf Nc33810d3dd054bdd9a1528d4816b3e4a
    34 Nf1b1de2361af4807915bd5dc58fb6aa5
    35 sg:journal.1040124
    36 schema:keywords C57BL/6N mice
    37 C57BL/6NTac
    38 CRISPR/
    39 Cas9 nickase
    40 Consortium Program
    41 DNA sequences
    42 HDR
    43 RNA
    44 RNA guides
    45 ResultsWe
    46 age
    47 alleles
    48 approach
    49 assessment
    50 auditory
    51 auditory phenotype
    52 bundles
    53 changes
    54 cochlea
    55 correction
    56 donor template
    57 electron microscopy
    58 genome
    59 guide
    60 guide RNA
    61 hair cell stereocilia bundles
    62 hearing loss
    63 hearing loss phenotype
    64 homology
    65 loss
    66 loss phenotype
    67 mice
    68 microscopy
    69 model
    70 mouse strains
    71 mutations
    72 nickase
    73 opportunities
    74 phenotype
    75 phenotypic assessment
    76 program
    77 progressive hearing loss
    78 progressive loss
    79 removal
    80 repair
    81 report
    82 rescue
    83 response testing
    84 sequence
    85 sequence changes
    86 sequencing
    87 sites
    88 specific DNA sequences
    89 stereocilia bundles
    90 strain-specific mutations
    91 strains
    92 study
    93 such mutations
    94 target
    95 targeting
    96 technology
    97 template
    98 testing
    99 weeks
    100 weeks of age
    101 whole-genome sequencing
    102 zygotes
    103 schema:name Correction of the auditory phenotype in C57BL/6N mice via CRISPR/Cas9-mediated homology directed repair
    104 schema:pagination 16
    105 schema:productId N0a2e8817f07f45b196bf8f292d6d0325
    106 N7d54ecb2c4854fb88e708f420430a016
    107 N7e2e5c850737461182462883c5435668
    108 schema:sameAs https://app.dimensions.ai/details/publication/pub.1045564567
    109 https://doi.org/10.1186/s13073-016-0273-4
    110 schema:sdDatePublished 2022-08-04T17:05
    111 schema:sdLicense https://scigraph.springernature.com/explorer/license/
    112 schema:sdPublisher N1db54f5788ef446da0adc7ae5c081d5c
    113 schema:url https://doi.org/10.1186/s13073-016-0273-4
    114 sgo:license sg:explorer/license/
    115 sgo:sdDataset articles
    116 rdf:type schema:ScholarlyArticle
    117 N03c7e03a65514e4db23c60322cfe9fc9 rdf:first sg:person.0632760571.83
    118 rdf:rest N74dbd241e5394aeb84af1d64a493f151
    119 N0a2e8817f07f45b196bf8f292d6d0325 schema:name dimensions_id
    120 schema:value pub.1045564567
    121 rdf:type schema:PropertyValue
    122 N0b8d118c20324bb18a4f1e3cb1f7a516 rdf:first sg:person.01047440473.56
    123 rdf:rest rdf:nil
    124 N168bb13c4cae43c287817007334fa5af schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
    125 schema:name Recombinational DNA Repair
    126 rdf:type schema:DefinedTerm
    127 N1db54f5788ef446da0adc7ae5c081d5c schema:name Springer Nature - SN SciGraph project
    128 rdf:type schema:Organization
    129 N3a9392da34b943b3baa0c6f77d1956f0 rdf:first sg:person.01337530626.28
    130 rdf:rest Nc9ac852361ed491abe7c76d1566db140
    131 N4455285f41b84de9babd631ab1c42bad schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
    132 schema:name Genetic Therapy
    133 rdf:type schema:DefinedTerm
    134 N461bbd2d98ba48349a86caca70a59ee4 rdf:first sg:person.01032576572.09
    135 rdf:rest N53cdcc7332204ee49b532d393120e4c1
    136 N4ff3f3b99b164bddb8397643dc6d2971 rdf:first sg:person.0601633627.85
    137 rdf:rest N03c7e03a65514e4db23c60322cfe9fc9
    138 N53cdcc7332204ee49b532d393120e4c1 rdf:first sg:person.012064305702.93
    139 rdf:rest N0b8d118c20324bb18a4f1e3cb1f7a516
    140 N5d045dc9a01c4343acf11b91dc46790a schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
    141 schema:name Animals
    142 rdf:type schema:DefinedTerm
    143 N611c89c6755e4952ba3c1cdc39b744fd schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
    144 schema:name Evoked Potentials, Auditory
    145 rdf:type schema:DefinedTerm
    146 N6d9bda0362d34954a3fea6f3cb031428 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
    147 schema:name Cadherins
    148 rdf:type schema:DefinedTerm
    149 N74dbd241e5394aeb84af1d64a493f151 rdf:first sg:person.01356107722.37
    150 rdf:rest Nfcc827e506e8444c9d4eba4ceefc25ea
    151 N7b23985906bd4019bc639552b70d1aaa rdf:first sg:person.016020736737.08
    152 rdf:rest N3a9392da34b943b3baa0c6f77d1956f0
    153 N7d54ecb2c4854fb88e708f420430a016 schema:name doi
    154 schema:value 10.1186/s13073-016-0273-4
    155 rdf:type schema:PropertyValue
    156 N7e2e5c850737461182462883c5435668 schema:name pubmed_id
    157 schema:value 26876963
    158 rdf:type schema:PropertyValue
    159 Na74bfd5c84ca4aa58fbe7a9308b7dbbf schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
    160 schema:name Stereocilia
    161 rdf:type schema:DefinedTerm
    162 Nb22a7c16c30f4696bd83e050ea6fcf87 rdf:first sg:person.0631215675.09
    163 rdf:rest Ndefa369d6a79409f8cbbbf8c09bcba86
    164 Nc33810d3dd054bdd9a1528d4816b3e4a schema:issueNumber 1
    165 rdf:type schema:PublicationIssue
    166 Nc9ac852361ed491abe7c76d1566db140 rdf:first sg:person.01244331330.82
    167 rdf:rest Nf780fe788f524d8aa5eef82f613f5875
    168 Nd2268ab1f7fe45639da25153c423fc82 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
    169 schema:name Hearing Loss
    170 rdf:type schema:DefinedTerm
    171 Nd83e1602b624437ba6ac0be43016b347 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
    172 schema:name Mice, Inbred C57BL
    173 rdf:type schema:DefinedTerm
    174 Ndefa369d6a79409f8cbbbf8c09bcba86 rdf:first sg:person.01100424027.43
    175 rdf:rest N7b23985906bd4019bc639552b70d1aaa
    176 Ne88b89c32c9a4376984347d11a3dede0 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
    177 schema:name Mice
    178 rdf:type schema:DefinedTerm
    179 Nf1b1de2361af4807915bd5dc58fb6aa5 schema:volumeNumber 8
    180 rdf:type schema:PublicationVolume
    181 Nf648736c5d5446a9a4d114fb8805bd24 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
    182 schema:name RNA, Guide
    183 rdf:type schema:DefinedTerm
    184 Nf780fe788f524d8aa5eef82f613f5875 rdf:first sg:person.01352202371.19
    185 rdf:rest N461bbd2d98ba48349a86caca70a59ee4
    186 Nf9d1c70f4cc74391bf674e313b5b7a22 schema:inDefinedTermSet https://www.nlm.nih.gov/mesh/
    187 schema:name CRISPR-Cas Systems
    188 rdf:type schema:DefinedTerm
    189 Nfcc827e506e8444c9d4eba4ceefc25ea rdf:first sg:person.01317506032.68
    190 rdf:rest Nb22a7c16c30f4696bd83e050ea6fcf87
    191 anzsrc-for:06 schema:inDefinedTermSet anzsrc-for:
    192 schema:name Biological Sciences
    193 rdf:type schema:DefinedTerm
    194 anzsrc-for:0604 schema:inDefinedTermSet anzsrc-for:
    195 schema:name Genetics
    196 rdf:type schema:DefinedTerm
    197 sg:grant.2752996 http://pending.schema.org/fundedItem sg:pub.10.1186/s13073-016-0273-4
    198 rdf:type schema:MonetaryGrant
    199 sg:grant.2756236 http://pending.schema.org/fundedItem sg:pub.10.1186/s13073-016-0273-4
    200 rdf:type schema:MonetaryGrant
    201 sg:grant.2782647 http://pending.schema.org/fundedItem sg:pub.10.1186/s13073-016-0273-4
    202 rdf:type schema:MonetaryGrant
    203 sg:grant.5498354 http://pending.schema.org/fundedItem sg:pub.10.1186/s13073-016-0273-4
    204 rdf:type schema:MonetaryGrant
    205 sg:grant.7440373 http://pending.schema.org/fundedItem sg:pub.10.1186/s13073-016-0273-4
    206 rdf:type schema:MonetaryGrant
    207 sg:grant.7921244 http://pending.schema.org/fundedItem sg:pub.10.1186/s13073-016-0273-4
    208 rdf:type schema:MonetaryGrant
    209 sg:journal.1040124 schema:issn 1756-994X
    210 schema:name Genome Medicine
    211 schema:publisher Springer Nature
    212 rdf:type schema:Periodical
    213 sg:person.01032576572.09 schema:affiliation grid-institutes:grid.420006.0
    214 schema:familyName Teboul
    215 schema:givenName Lydia
    216 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01032576572.09
    217 rdf:type schema:Person
    218 sg:person.01047440473.56 schema:affiliation grid-institutes:grid.420006.0
    219 schema:familyName Bowl
    220 schema:givenName Michael R.
    221 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01047440473.56
    222 rdf:type schema:Person
    223 sg:person.01100424027.43 schema:affiliation grid-institutes:grid.420006.0
    224 schema:familyName Hutchison
    225 schema:givenName Marie
    226 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01100424027.43
    227 rdf:type schema:Person
    228 sg:person.012064305702.93 schema:affiliation grid-institutes:grid.420006.0
    229 schema:familyName Brown
    230 schema:givenName Steve D. M.
    231 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.012064305702.93
    232 rdf:type schema:Person
    233 sg:person.01244331330.82 schema:affiliation grid-institutes:grid.420006.0
    234 schema:familyName Wells
    235 schema:givenName Sara
    236 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01244331330.82
    237 rdf:type schema:Person
    238 sg:person.01317506032.68 schema:affiliation grid-institutes:grid.420006.0
    239 schema:familyName Aguilar
    240 schema:givenName Carlos
    241 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01317506032.68
    242 rdf:type schema:Person
    243 sg:person.01337530626.28 schema:affiliation grid-institutes:grid.420006.0
    244 schema:familyName Mallon
    245 schema:givenName Ann-Marie
    246 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01337530626.28
    247 rdf:type schema:Person
    248 sg:person.01352202371.19 schema:affiliation grid-institutes:grid.420006.0
    249 schema:familyName Simon
    250 schema:givenName Michelle M.
    251 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01352202371.19
    252 rdf:type schema:Person
    253 sg:person.01356107722.37 schema:affiliation grid-institutes:grid.420006.0
    254 schema:familyName Kumar
    255 schema:givenName Saumya
    256 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.01356107722.37
    257 rdf:type schema:Person
    258 sg:person.016020736737.08 schema:affiliation grid-institutes:grid.420006.0
    259 schema:familyName Parker
    260 schema:givenName Andrew
    261 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.016020736737.08
    262 rdf:type schema:Person
    263 sg:person.0601633627.85 schema:affiliation grid-institutes:grid.420006.0
    264 schema:familyName Mianné
    265 schema:givenName Joffrey
    266 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0601633627.85
    267 rdf:type schema:Person
    268 sg:person.0631215675.09 schema:affiliation grid-institutes:grid.420006.0
    269 schema:familyName Codner
    270 schema:givenName Gemma
    271 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0631215675.09
    272 rdf:type schema:Person
    273 sg:person.0632760571.83 schema:affiliation grid-institutes:grid.420006.0
    274 schema:familyName Chessum
    275 schema:givenName Lauren
    276 schema:sameAs https://app.dimensions.ai/discover/publication?and_facet_researcher=ur.0632760571.83
    277 rdf:type schema:Person
    278 sg:pub.10.1007/978-1-60327-019-9_8 schema:sameAs https://app.dimensions.ai/details/publication/pub.1016698235
    279 https://doi.org/10.1007/978-1-60327-019-9_8
    280 rdf:type schema:CreativeWork
    281 sg:pub.10.1007/s00335-012-9427-x schema:sameAs https://app.dimensions.ai/details/publication/pub.1009915802
    282 https://doi.org/10.1007/s00335-012-9427-x
    283 rdf:type schema:CreativeWork
    284 sg:pub.10.1007/s101620010075 schema:sameAs https://app.dimensions.ai/details/publication/pub.1038584115
    285 https://doi.org/10.1007/s101620010075
    286 rdf:type schema:CreativeWork
    287 sg:pub.10.1038/ejhg.2009.234 schema:sameAs https://app.dimensions.ai/details/publication/pub.1036164743
    288 https://doi.org/10.1038/ejhg.2009.234
    289 rdf:type schema:CreativeWork
    290 sg:pub.10.1038/ejhg.2014.56 schema:sameAs https://app.dimensions.ai/details/publication/pub.1031381434
    291 https://doi.org/10.1038/ejhg.2014.56
    292 rdf:type schema:CreativeWork
    293 sg:pub.10.1038/nature10413 schema:sameAs https://app.dimensions.ai/details/publication/pub.1041325874
    294 https://doi.org/10.1038/nature10413
    295 rdf:type schema:CreativeWork
    296 sg:pub.10.1038/nbt.2623 schema:sameAs https://app.dimensions.ai/details/publication/pub.1009511624
    297 https://doi.org/10.1038/nbt.2623
    298 rdf:type schema:CreativeWork
    299 sg:pub.10.1038/nbt.2647 schema:sameAs https://app.dimensions.ai/details/publication/pub.1012543463
    300 https://doi.org/10.1038/nbt.2647
    301 rdf:type schema:CreativeWork
    302 sg:pub.10.1038/nmeth.2857 schema:sameAs https://app.dimensions.ai/details/publication/pub.1035757498
    303 https://doi.org/10.1038/nmeth.2857
    304 rdf:type schema:CreativeWork
    305 sg:pub.10.1038/nprot.2009.204 schema:sameAs https://app.dimensions.ai/details/publication/pub.1039038752
    306 https://doi.org/10.1038/nprot.2009.204
    307 rdf:type schema:CreativeWork
    308 sg:pub.10.1038/nprot.2013.143 schema:sameAs https://app.dimensions.ai/details/publication/pub.1016534425
    309 https://doi.org/10.1038/nprot.2013.143
    310 rdf:type schema:CreativeWork
    311 sg:pub.10.1038/nrg2309 schema:sameAs https://app.dimensions.ai/details/publication/pub.1001777660
    312 https://doi.org/10.1038/nrg2309
    313 rdf:type schema:CreativeWork
    314 sg:pub.10.1038/srep05396 schema:sameAs https://app.dimensions.ai/details/publication/pub.1011163022
    315 https://doi.org/10.1038/srep05396
    316 rdf:type schema:CreativeWork
    317 grid-institutes:grid.420006.0 schema:alternateName Mammalian Genetics Unit, MRC Harwell, Harwell, OX11 0RD, Oxford, UK
    318 Mary Lyon Centre, MRC Harwell, Harwell, OX11 0RD, Oxford, UK
    319 schema:name Mammalian Genetics Unit, MRC Harwell, Harwell, OX11 0RD, Oxford, UK
    320 Mary Lyon Centre, MRC Harwell, Harwell, OX11 0RD, Oxford, UK
    321 rdf:type schema:Organization
     




    Preview window. Press ESC to close (or click here)


    ...