Journal article Open Access

Efficient Generation and Correction of Mutations in Human iPS Cells Utilizing mRNAs of CRISPR Base Editors and Prime Editors

Sürün, Duran; Schneider, Aksana; Mircetic, Jovan; Neumann, Katrin; Lansing, Felix; Paszkowski-Rogacz, Maciej; Hänchen, Vanessa; Lee-Kirsch, Min Ae; Buchholz, Frank


JSON-LD (schema.org) Export

{
  "description": "<p><strong>Abstract</strong></p>\n\n<p>In contrast to CRISPR/Cas9 nucleases, CRISPR base editors (BE) and prime editors (PE) enable predefined nucleotide exchanges in genomic sequences without generating DNA double strand breaks. Here, we employed BE and PE mRNAs in conjunction with chemically synthesized sgRNAs and pegRNAs for efficient editing of human induced pluripotent stem cells (iPSC). Whereas we were unable to correct a disease-causing mutation in patient derived iPSCs using a CRISPR/Cas9 nuclease approach, we corrected the mutation back to wild type with high efficiency utilizing an adenine BE. We also used adenine and cytosine BEs to introduce nine different cancer associated&nbsp;TP53&nbsp;mutations into human iPSCs with up to 90% efficiency, generating a panel of cell lines to investigate the biology of these mutations in an isogenic background. Finally, we pioneered the use of prime editing in human iPSCs, opening this important cell type for the precise modification of nucleotides not addressable by BEs and to multiple nucleotide exchanges. These approaches eliminate the necessity of deriving disease specific iPSCs from human donors and allows the comparison of different disease-causing mutations in isogenic genetic backgrounds.</p>", 
  "license": "https://creativecommons.org/licenses/by/4.0/legalcode", 
  "creator": [
    {
      "affiliation": "Medical Systems Biology, Medical Faculty and University Hospital Carl Gustav Carus, TU Dresden, 01307 Dresden, Germany", 
      "@type": "Person", 
      "name": "S\u00fcr\u00fcn,\u00a0Duran"
    }, 
    {
      "affiliation": "Medical Systems Biology, Medical Faculty and University Hospital Carl Gustav Carus, TU Dresden, 01307 Dresden, Germany", 
      "@type": "Person", 
      "name": "Schneider,\u00a0Aksana"
    }, 
    {
      "affiliation": "Medical Systems Biology, Medical Faculty and University Hospital Carl Gustav Carus, TU Dresden, 01307 Dresden, Germany - Mildred Scheel Early Career Center, National Center for Tumor Diseases Dresden (NCT/UCC), Medical Faculty and University Hospital Carl Gustav Carus, TU Dresden, 01307 Dresden, Germany", 
      "@type": "Person", 
      "name": "Mircetic,\u00a0Jovan"
    }, 
    {
      "affiliation": "Stem Cell Engineering Facility, Center for Molecular and Cellular Bioengineering (CMCB), TU Dresden, 01307 Dresden, Germany", 
      "@type": "Person", 
      "name": "Neumann,\u00a0Katrin"
    }, 
    {
      "affiliation": "Medical Systems Biology, Medical Faculty and University Hospital Carl Gustav Carus, TU Dresden, 01307 Dresden, Germany", 
      "@type": "Person", 
      "name": "Lansing,\u00a0Felix"
    }, 
    {
      "affiliation": "Medical Systems Biology, Medical Faculty and University Hospital Carl Gustav Carus, TU Dresden, 01307 Dresden, Germany", 
      "@type": "Person", 
      "name": "Paszkowski-Rogacz,\u00a0Maciej"
    }, 
    {
      "affiliation": "Department of Pediatrics, Medical Faculty and University Hospital Carl Gustav Carus, TU Dresden, 01307 Dresden, Germany", 
      "@type": "Person", 
      "name": "H\u00e4nchen,\u00a0Vanessa"
    }, 
    {
      "affiliation": "Department of Pediatrics, Medical Faculty and University Hospital Carl Gustav Carus, TU Dresden, 01307 Dresden, Germany", 
      "@type": "Person", 
      "name": "Lee-Kirsch,\u00a0Min Ae"
    }, 
    {
      "affiliation": "Medical Systems Biology, Medical Faculty and University Hospital Carl Gustav Carus, TU Dresden, 01307 Dresden, Germany", 
      "@type": "Person", 
      "name": "Buchholz,\u00a0Frank"
    }
  ], 
  "headline": "Efficient Generation and Correction of Mutations in Human iPS Cells Utilizing mRNAs of CRISPR Base Editors and Prime Editors", 
  "image": "https://zenodo.org/static/img/logos/zenodo-gradient-round.svg", 
  "datePublished": "2020-05-06", 
  "url": "https://zenodo.org/record/5751965", 
  "@context": "https://schema.org/", 
  "identifier": "https://doi.org/10.3390/genes11050511", 
  "@id": "https://doi.org/10.3390/genes11050511", 
  "@type": "ScholarlyArticle", 
  "name": "Efficient Generation and Correction of Mutations in Human iPS Cells Utilizing mRNAs of CRISPR Base Editors and Prime Editors"
}
34
18
views
downloads
Views 34
Downloads 18
Data volume 45.4 MB
Unique views 32
Unique downloads 18

Share

Cite as