ArticleEukaryotic cell2005
Nuclear-gene targeting by using single-stranded DNA avoids illegitimate DNA integration in Chlamydomonas reinhardtii.
Article in Eukaryotic cell, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 37 papers.
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Who cites it
37 citing papers in PubMed, 88 citations in OpenAlex.
- Epigenetic Regulation of Salt Stress Responses in Rice: Mechanisms and Prospects for Enhancing Tolerance.Epigenomes · 2025Review
- Efficient non-viral immune cell engineering using circular single-stranded DNA-mediated genomic integration.Nature biotechnology · 2025Article
- The S862C amino acid change in CpMrr1 confers fluconazole resistance inJAC-antimicrobial resistance · 2025Article
- Biomanufacturing of γ-linolenic acid-enriched galactosyldiacylglycerols: Challenges in microalgae and potential in oleaginous yeasts.Synthetic and systems biotechnology · 2023Review
- Improved selection of zebrafish CRISPR editing by early next-generation sequencing based genotyping.Scientific reports · 2023Article
- Two stones, one bird: co-targeting facilitates precise gene editing of non-selectable genes in Chlamydomonas.Plant physiology · 2022Article
- Optimized two-step electroporation process to achieve efficient nonviral-mediated gene insertion into primary T cells.FEBS open bio · 2022Article
- Mechanistic and genetic basis of single-strand templated repair at Cas12a-induced DNA breaks in Chlamydomonas reinhardtii.Nature communications · 2021Article
- Architecture and evolution of subtelomeres in the unicellular green alga Chlamydomonas reinhardtii.Nucleic acids research · 2021Article
- Comparative genomics of Chlamydomonas.The Plant cell · 2021Article
- Review
- Article
- Genome Editing by CRISPR-Cas: A Game Change in the Genetic Manipulation ofLife (Basel, Switzerland) · 2020Review
- CRISPR based targeted genome editing of Chlamydomonas reinhardtii using programmed Cas9-gRNA ribonucleoprotein.Molecular biology reports · 2020Article
- A cell cycle-dependent CRISPR-Cas9 activation system based on an anti-CRISPR protein shows improved genome editing accuracy.Communications biology · 2020Article
- Efficient In Vivo Introduction of Point Mutations Using ssODN and a Co-CRISPR Approach.Biological procedures online · 2020Article
- Manipulation of the microalgal chloroplast by genetic engineering for biotechnological utilization as a green biofactory.World journal of microbiology & biotechnology · 2018Review
- CRISPR/Cas9-mediated homology-directed repair by ssODNs in zebrafish induces complex mutational patterns resulting from genomic integration of repair-template fragments.Disease models & mechanisms · 2018Article
- Efficient targeted DNA editing and replacement inProceedings of the National Academy of Sciences of the United States of America · 2017Article
- ClippingThe Plant cell · 2017Article
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Homologous DNA recombination (HR) allows the deletion (knockout), repair (rescuing), and modification of a selected gene, thereby rendering a functional analysis of the gene product possible. However, targeting of nuclear genes has been an inefficient process in most eukaryotes, including algae, plants, and animals, due to the dominance of integration of the applied DNA into nonhomologous regions of the genome. We have shown for the green alga Chlamydomonas reinhardtii by repairing a previously introduced truncated aminoglycoside 3'-phosphotransferase gene, aphVIII, that single-stranded DNA can recombine with a homologous endogenous DNA region of interest. Nonhomologous DNA integration appeared to be more than 100-fold reduced compared with the use of double-stranded DNA, thus allowing isolation of the homologous recombinants. We propose that this method will be applicable to direct targeting of nuclear C. reinhardtii genes.
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