Circular synthesized CRISPR/Cas gRNAs for functional interrogations in the coding and noncoding genome Article Swipe
Martin Wegner
,
Valentina Diehl
,
Verena Bittl
,
Rahel de Bruyn
,
Svenja Wiechmann
,
Yves Matthess
,
Marie Hebel
,
Michael G.B. Hayes
,
Simone Schaubeck
,
Christopher Benner
,
Sven Heinz
,
Anja Bremm
,
Ivan Đikić
,
Andreas Ernst
,
Manuel Kaulich
·
YOU?
·
· 2019
· Open Access
·
· DOI: https://doi.org/10.7554/elife.42549
YOU?
·
· 2019
· Open Access
·
· DOI: https://doi.org/10.7554/elife.42549
Current technologies used to generate CRISPR/Cas gene perturbation reagents are labor intense and require multiple ligation and cloning steps. Furthermore, increasing gRNA sequence diversity negatively affects gRNA distribution, leading to libraries of heterogeneous quality. Here, we present a rapid and cloning-free mutagenesis technology that can efficiently generate covalently-closed-circular-synthesized (3Cs) CRISPR/Cas gRNA reagents and that uncouples sequence diversity from sequence distribution. We demonstrate the fidelity and performance of 3Cs reagents by tailored targeting of all human deubiquitinating enzymes (DUBs) and identify their essentiality for cell fitness. To explore high-content screening, we aimed to generate the largest up-to-date gRNA library that can be used to interrogate the coding and noncoding human genome and simultaneously to identify genes, predicted promoter flanking regions, transcription factors and CTCF binding sites that are linked to doxorubicin resistance. Our 3Cs technology enables fast and robust generation of bias-free gene perturbation libraries with yet unmatched diversities and should be considered an alternative to established technologies.
Related Topics
Metadata
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.7554/elife.42549
- OA Status
- gold
- Cited By
- 43
- References
- 66
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W2919380816
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W2919380816Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.7554/elife.42549Digital Object Identifier
- Title
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Circular synthesized CRISPR/Cas gRNAs for functional interrogations in the coding and noncoding genomeWork title
- Type
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articleOpenAlex work type
- Language
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enPrimary language
- Publication year
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2019Year of publication
- Publication date
-
2019-03-06Full publication date if available
- Authors
-
Martin Wegner, Valentina Diehl, Verena Bittl, Rahel de Bruyn, Svenja Wiechmann, Yves Matthess, Marie Hebel, Michael G.B. Hayes, Simone Schaubeck, Christopher Benner, Sven Heinz, Anja Bremm, Ivan Đikić, Andreas Ernst, Manuel KaulichList of authors in order
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https://doi.org/10.7554/elife.42549Publisher landing page
- Open access
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YesWhether a free full text is available
- OA status
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goldOpen access status per OpenAlex
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https://doi.org/10.7554/elife.42549Direct OA link when available
- Concepts
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CRISPR, Guide RNA, Computational biology, Biology, Gene, Genome, Genetics, Genome editingTop concepts (fields/topics) attached by OpenAlex
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-
43Total citation count in OpenAlex
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2025: 2, 2024: 5, 2023: 13, 2022: 6, 2021: 6Per-year citation counts (last 5 years)
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10Other works algorithmically related by OpenAlex
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