Efficient Generation and Correction of Mutations in Human iPS Cells Utilizing mRNAs of CRISPR Base Editors and Prime Editors Article Swipe
Duran Sürün
,
Aksana Schneider
,
Jovan Mircetic
,
Katrin Neumann
,
Felix Lansing
,
Maciej Paszkowski‐Rogacz
,
Vanessa Hänchen
,
Min Ae Lee‐Kirsch
,
Frank Buchholz
·
YOU?
·
· 2020
· Open Access
·
· DOI: https://doi.org/10.3390/genes11050511
YOU?
·
· 2020
· Open Access
·
· DOI: https://doi.org/10.3390/genes11050511
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 TP53 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.
Related Topics
Concepts
CRISPR
Induced pluripotent stem cell
Cas9
Genome editing
Nuclease
Biology
Mutation
Computational biology
Genetics
DNA
Nucleotide
Gene
Embryonic stem cell
Metadata
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.3390/genes11050511
- OA Status
- gold
- Cited By
- 121
- References
- 55
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W3023181423
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W3023181423Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.3390/genes11050511Digital Object Identifier
- Title
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Efficient Generation and Correction of Mutations in Human iPS Cells Utilizing mRNAs of CRISPR Base Editors and Prime EditorsWork title
- Type
-
articleOpenAlex work type
- Language
-
enPrimary language
- Publication year
-
2020Year of publication
- Publication date
-
2020-05-06Full publication date if available
- Authors
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Duran Sürün, Aksana Schneider, Jovan Mircetic, Katrin Neumann, Felix Lansing, Maciej Paszkowski‐Rogacz, Vanessa Hänchen, Min Ae Lee‐Kirsch, Frank BuchholzList of authors in order
- Landing page
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https://doi.org/10.3390/genes11050511Publisher landing page
- Open access
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YesWhether a free full text is available
- OA status
-
goldOpen access status per OpenAlex
- OA URL
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https://doi.org/10.3390/genes11050511Direct OA link when available
- Concepts
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CRISPR, Induced pluripotent stem cell, Cas9, Genome editing, Nuclease, Biology, Mutation, Computational biology, Genetics, DNA, Nucleotide, Gene, Embryonic stem cellTop concepts (fields/topics) attached by OpenAlex
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121Total citation count in OpenAlex
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2025: 18, 2024: 18, 2023: 18, 2022: 27, 2021: 32Per-year citation counts (last 5 years)
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55Number of works referenced by this work
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-
10Other works algorithmically related by OpenAlex
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| primary_topic.display_name | CRISPR and Genetic Engineering |
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