CRIMP: a CRISPR/Cas9 insertional mutagenesis protocol and toolkit Article Swipe
Lee B. Miles
,
Vanessa Calcinotto
,
Sara Oveissi
,
Rita J. Serrano
,
C. Sonntag
,
Orlen Mulia
,
Clara Lee
,
Robert J. Bryson‐Richardson
·
YOU?
·
· 2024
· Open Access
·
· DOI: https://doi.org/10.1038/s41467-024-49341-7
YOU?
·
· 2024
· Open Access
·
· DOI: https://doi.org/10.1038/s41467-024-49341-7
Site-directed insertion is a powerful approach for generating mutant alleles, but low efficiency and the need for customisation for each target has limited its application. To overcome this, we developed a highly efficient targeted insertional mutagenesis system, CRIMP, and an associated plasmid toolkit, CRIMPkit, that disrupts native gene expression by inducing complete transcriptional termination, generating null mutant alleles without inducing genetic compensation. The protocol results in a high frequency of integration events and can generate very early targeted insertions, during the first cell division, producing embryos with expression in one or both halves of the body plan. Fluorescent readout of integration events facilitates selection of successfully mutagenized fish and, subsequently, visual identification of heterozygous and mutant animals. Together, these advances greatly improve the efficacy of generating and studying mutant lines. The CRIMPkit contains 24 ready-to-use plasmid vectors to allow easy and complete mutagenesis of any gene in any reading frame without requiring custom sequences, modification, or subcloning.
Related Topics
Concepts
CRISPR
Computational biology
Insertional mutagenesis
Crimp
Biology
Mutagenesis
Protocol (science)
Computer science
Genetics
Mutation
Medicine
Gene
Genome
Chemistry
Pathology
Physical chemistry
Alternative medicine
Metadata
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1038/s41467-024-49341-7
- https://www.nature.com/articles/s41467-024-49341-7.pdf
- OA Status
- gold
- Cited By
- 5
- References
- 46
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4399584625
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W4399584625Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.1038/s41467-024-49341-7Digital Object Identifier
- Title
-
CRIMP: a CRISPR/Cas9 insertional mutagenesis protocol and toolkitWork title
- Type
-
articleOpenAlex work type
- Language
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enPrimary language
- Publication year
-
2024Year of publication
- Publication date
-
2024-06-12Full publication date if available
- Authors
-
Lee B. Miles, Vanessa Calcinotto, Sara Oveissi, Rita J. Serrano, C. Sonntag, Orlen Mulia, Clara Lee, Robert J. Bryson‐RichardsonList of authors in order
- Landing page
-
https://doi.org/10.1038/s41467-024-49341-7Publisher landing page
- PDF URL
-
https://www.nature.com/articles/s41467-024-49341-7.pdfDirect link to full text PDF
- Open access
-
YesWhether a free full text is available
- OA status
-
goldOpen access status per OpenAlex
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https://www.nature.com/articles/s41467-024-49341-7.pdfDirect OA link when available
- Concepts
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CRISPR, Computational biology, Insertional mutagenesis, Crimp, Biology, Mutagenesis, Protocol (science), Computer science, Genetics, Mutation, Medicine, Gene, Genome, Chemistry, Pathology, Physical chemistry, Alternative medicineTop concepts (fields/topics) attached by OpenAlex
- Cited by
-
5Total citation count in OpenAlex
- Citations by year (recent)
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2025: 3, 2024: 1, 2023: 1Per-year citation counts (last 5 years)
- References (count)
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46Number of works referenced by this work
- Related works (count)
-
10Other works algorithmically related by OpenAlex
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