CRISPR-STOP: gene silencing through base-editing-induced nonsense mutations
Abstract
CRISPR-Cas9-induced DNA damage may have deleterious effects at high-copy-number genomic regions. Here, we use CRISPR base editors to knock out genes by changing single nucleotides to create stop codons. We show that the CRISPR-STOP method is an efficient and less deleterious alternative to wild-type Cas9 for gene-knockout studies. Early stop codons can be introduced in ∼17,000 human genes. CRISPR-STOP-mediated targeted screening demonstrates comparable efficiency to WT Cas9, which indicates the suitability of our approach for genome-wide functional screenings.
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Base editing can be repurposed as a gene loss of function tool by creating a stop codon or a gene knockdown by creating an upstream open reading frame (uORF).
“It can also be repurposed as a gene loss of function tool by creating a stop codon or a gene knockdown by creating an upstream open reading frame (uORF) (Billon et al., 2017;Kuscu et al., 2017;Xiong et al., 2022).”
From Genetic resources and precise gene editing for targeted improvement of barley abiotic stress tolerance (2023) · cites this paper as Kuscu et al., 2017
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Research Progress on Cloning and Function of Xa Genes Against Rice Bacterial Blight
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“Initial screens with base editor technology in mammalian cells have introduced stop codons (Kuscu et al., 2017) or tiled all possible sgRNAs across a small number of genes (Jun et al., 2020; Kweon et al., 2019), and such screens have recently been applied at scale in yeast (Despres et al., 2020).”
Programmable Base Editing of the Sheep Genome Revealed No Genome-Wide Off-Target Mutations
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CRISPR-Based Genetic Manipulation of Candida Species: Historical Perspectives and Current Approaches
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Reprogramming Acetogenic Bacteria with CRISPR-Targeted Base Editing via Deamination
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- Authors
- Kuscu, C., Parlak, M., Tufan, T., Yang, J., Szlachta, K., Wei, X., Mammadov, R., & Adli, M.
- Affiliations
- University of Virginia (US)
- Corresponding author
- Adli, Mazhar
- ORCID
- Venue
- Nature Methods
- Published
- 2017-06-05
- Volume
- 14
- Issue
- 7
- Pages
- 710-712
- ISSN
- 1548-7091 (ISSN-L)
- 1548-7105
- OpenAlex
- W2621848900
- Cited by
- 344
- References
- 21
- Citations by year
- 2026: 21 · 2025: 28 · 2024: 33 · 2023: 43 · 2022: 35 · 2021: 45 · 2020: 44 · 2019: 42 · 2018: 44 · 2017: 9
- FWCI
- 15.52
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Methods (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Keywords
- base editing, nonsense mutation, gene knockout, CRISPR/Cas9, targeted mutagenesis, single-nucleotide editing
- MeSH terms
- Gene Expression Regulation, Genetic Vectors, Humans, Plasmids, Codon, Terminator, Codon, Nonsense, Gene Targeting, Gene Silencing, HEK293 Cells, Clustered Regularly Interspaced Short Palindromic Repeats
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