ArticleBiological procedures online2020
Efficient In Vivo Introduction of Point Mutations Using ssODN and a Co-CRISPR Approach.
Article in Biological procedures online, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed.
- Coordinated active repression operates via transcription factor cooperativity and multiple inactive promoter states in a developing organism.Nature communications · 2025Article
- From promoter motif to cardiac function: a single DPE motif affects transcription regulation and organ function in vivo.Development (Cambridge, England) · 2024Article
- Engineering and Expression Strategies for Optimization of L-Asparaginase Development and Production.International journal of molecular sciences · 2023Review
- CRISPR/Cas9-mediated knock-in inGene and genome editing · 2023Article
- Precise CRISPR-Cas9-mediated mutation of a membrane trafficking domain in theCurrent research in physiology · 2023Article
- Article
- Improving one-step scarless genome editing inBiology methods & protocols · 2022Article
- Precise genome engineering inProceedings of the National Academy of Sciences of the United States of America · 2021Article
- The Core Promoter Is a Regulatory Hub for Developmental Gene Expression.Frontiers in cell and developmental biology · 2021Review
Corrections and comments
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundThe generation of point mutations is a major tool for evaluating the roles of specific nucleotides or amino acids within the regulatory or functional landscape. However, examination of these mutations in vivo requires the generation of animals carrying only the relevant point mutations at the endogenous genomic loci, which is technically challenging. The CRISPR-Cas9 based genome editing greatly facilitates the generation of such genetically modified animals; however, most of the described methods use double-strand DNA (dsDNA) as the donor template. The dsDNA plasmids frequently undergo undesired integration events into the targeted genomic locus. The use of a single-strand oligodeoxynucleotide (ssODN) as the donor template prevents this complication and is therefore the preferred choice for introducing point mutations, as well as short sequences such as protein tags.
resultsWe successfully applied the CRISPR-based
conclusionsThe described approach has facilitated the successful generation of point mutations in two different chromosomes, by two different labs. Distinct phenotypes associated with the newly-generated genotype were identified, thus exemplifying the importance of investigating the in vivo role of specific nucleotides. In addition, detailed guidelines, recommendations and crossing schemes are provided in order to support the generation of additional genetically modified animals by the scientific community.
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Registered trials
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