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Multiplex Conditional Mutagenesis Using Transgenic Expression of Cas9 and sgRNAs

Linlin Yin, Lisette A. Maddison, View ORCID ProfileMingyu Li, Nergis Kara, View ORCID ProfileMatthew C. LaFave, View ORCID ProfileGaurav K. Varshney, View ORCID ProfileShawn M. Burgess, View ORCID ProfileJames G. Patton and View ORCID ProfileWenbiao Chen
Genetics June 1, 2015 vol. 200 no. 2 431-441; https://doi.org/10.1534/genetics.115.176917
Linlin Yin
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee 37232
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Lisette A. Maddison
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee 37232
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Mingyu Li
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee 37232
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  • Search for this author on this site
  • ORCID record for Mingyu Li
Nergis Kara
Department of Biological Science, Vanderbilt University, Nashville, Tennessee 37240
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Matthew C. LaFave
Translational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892
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  • Search for this author on this site
  • ORCID record for Matthew C. LaFave
Gaurav K. Varshney
Translational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892
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  • ORCID record for Gaurav K. Varshney
Shawn M. Burgess
Translational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892
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  • ORCID record for Shawn M. Burgess
James G. Patton
Department of Biological Science, Vanderbilt University, Nashville, Tennessee 37240
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  • ORCID record for James G. Patton
Wenbiao Chen
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee 37232
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  • ORCID record for Wenbiao Chen
  • For correspondence: wenbiao.chen@vanderbilt.edu
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Data supplements

  • Files in this data supplement:

    • Supporting Information: Figure S1, Tables S1-S5, and File S1 (PDF, 430 KB)
    • Figure S1: Alignment of top 5 alleles of the 3 loci in Tg(actb2:cas9; (U6x:sgRNA(insra/b) fish. (PDF, 707 KB)
    • Table S1: List of primers use in the study. (.xlsx, 11 KB)
    • Table S2: Mutation rates at the three loci calculated based on high throughput sequencing data. (.xlsx, 55 KB)
    • Table S3: Alleles and read counts of insra locus identified in the 17 double carriers. "D" denotes deletion. "I" denotes insertion. "C" denotes deletion and insertion. The first number indicates the position at which InDel occurs. The second number is the number of nucleotides of insertion or deletion in the allele. (.xlsx, 404 KB)
    • Table S4: Alleles and read counts of insrb locus identified in the 17 double carriers. (.xlsx, 264 KB)
    • Table S5: Alleles and read counts of tyr locus identified in the 17 double carriers. (.xlsx, 75 KB)
    • File S1: Supplemental Methods (PDF, 667 KB)
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PUBLICATION INFORMATION

Volume 200 Issue 2, June 2015

Genetics: 200 (2)

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  • Methods, Technology, & Resources

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Multiplex Conditional Mutagenesis Using Transgenic Expression of Cas9 and sgRNAs

Linlin Yin, Lisette A. Maddison, View ORCID ProfileMingyu Li, Nergis Kara, View ORCID ProfileMatthew C. LaFave, View ORCID ProfileGaurav K. Varshney, View ORCID ProfileShawn M. Burgess, View ORCID ProfileJames G. Patton and View ORCID ProfileWenbiao Chen
Genetics June 1, 2015 vol. 200 no. 2 431-441; https://doi.org/10.1534/genetics.115.176917
Linlin Yin
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee 37232
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Lisette A. Maddison
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee 37232
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Mingyu Li
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee 37232
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Mingyu Li
Nergis Kara
Department of Biological Science, Vanderbilt University, Nashville, Tennessee 37240
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Matthew C. LaFave
Translational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Matthew C. LaFave
Gaurav K. Varshney
Translational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Gaurav K. Varshney
Shawn M. Burgess
Translational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Shawn M. Burgess
James G. Patton
Department of Biological Science, Vanderbilt University, Nashville, Tennessee 37240
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for James G. Patton
Wenbiao Chen
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee 37232
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Wenbiao Chen
  • For correspondence: wenbiao.chen@vanderbilt.edu
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Citation

Multiplex Conditional Mutagenesis Using Transgenic Expression of Cas9 and sgRNAs

Linlin Yin, Lisette A. Maddison, View ORCID ProfileMingyu Li, Nergis Kara, View ORCID ProfileMatthew C. LaFave, View ORCID ProfileGaurav K. Varshney, View ORCID ProfileShawn M. Burgess, View ORCID ProfileJames G. Patton and View ORCID ProfileWenbiao Chen
Genetics June 1, 2015 vol. 200 no. 2 431-441; https://doi.org/10.1534/genetics.115.176917
Linlin Yin
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee 37232
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Lisette A. Maddison
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee 37232
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Mingyu Li
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee 37232
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Mingyu Li
Nergis Kara
Department of Biological Science, Vanderbilt University, Nashville, Tennessee 37240
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Matthew C. LaFave
Translational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Matthew C. LaFave
Gaurav K. Varshney
Translational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Gaurav K. Varshney
Shawn M. Burgess
Translational and Functional Genomics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Shawn M. Burgess
James G. Patton
Department of Biological Science, Vanderbilt University, Nashville, Tennessee 37240
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for James G. Patton
Wenbiao Chen
Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee 37232
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Wenbiao Chen
  • For correspondence: wenbiao.chen@vanderbilt.edu

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Methods, Technology, and Resources

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  • NATF (Native and Tissue-Specific Fluorescence): A Strategy for Bright, Tissue-Specific GFP Labeling of Native Proteins in Caenorhabditis elegans
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Show more Methods, Technology, and Resources
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  • Figures & Data
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