The DOI matches its Crossref record; 1 of the 1 references it cites were found in a registry or an index.
High efficiency mutagenesis, repair, and engineering of chromosomal DNA using single-stranded oligonucleotides
Open accessAbstract
Homologous DNA recombination is a fundamental, regenerative process within living organisms. However, in most organisms, homologous recombination is a rare event, requiring a complex set of reactions and extensive homology. We demonstrate in this paper that Beta protein of phage λ generates recombinants in chromosomal DNA by using synthetic single-stranded DNAs (ssDNA) as short as 30 bases long. This ssDNA recombination can be used to mutagenize or repair the chromosome with efficiencies that generate up to 6% recombinants among treated cells. Mechanistically, it appears that Beta protein, a Rad52-like protein, binds and anneals the ssDNA donor to a complementary single-strand near the DNA replication fork to generate the recombinant. This type of homologous recombination with ssDNA provides new avenues for studying and modifying genomes ranging from bacterial pathogens to eukaryotes. Beta protein and ssDNA may prove generally applicable for repairing DNA in many organisms.
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Cites
The DNA damage response: putting checkpoints in perspective
Zhou, B.-B. S., & Elledge, S. J. (2000) · Nature · doi:10.1038/35044005 · Cited by 3,213
Cited by
Programming cells by multiplex genome engineering and accelerated evolution
Wang, H. H., Isaacs, F. J., Carr, P. A., Sun, Z. Z., Xu, G., Forest, C. R., & Church, G. M. (2009) · Nature · doi:10.1038/nature08187 · Cited by 1,529
“Oligonucleotides used in the MAGE process contained the following modifications: (1) 30–110 bp in length for optimization experiments; (2) different numbers of phosphorothioated bases; and (3) degenerate nucleotides as described elsewhere in this paper.”
BackgroundMethodRecombineering: a homologous recombination-based method of genetic engineering
Sharan, S. K., Thomason, L. C., Kuznetsov, S. G., & Court, D. L. (2009) · Nature Protocols · doi:10.1038/nprot.2008.227 · Cited by 785
Self-citationWu, T., Malinverni, J., Ruiz, N., Kim, S., Silhavy, T. J., & Kahne, D. (2005) · Cell · doi:10.1016/j.cell.2005.02.015 · Cited by 769
Directed Evolution: Methodologies and Applications
Wang, Y., Xue, P., Cao, M., Yu, T., Lane, S. T., & Zhao, H. (2021) · Chemical Reviews · doi:10.1021/acs.chemrev.1c00260 · Cited by 719
En Passant Mutagenesis: A Two Step Markerless Red Recombination System
Tischer, B. K., Smith, G. A., & Osterrieder, N. (2010) · Methods in Molecular Biology · doi:10.1007/978-1-60761-652-8_30 · Cited by 599
Metadata
- Authors
- Ellis, H. M., Yu, D., DiTizio, T., & Court, D. L.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 2001-05-29
- Volume
- 98
- Issue
- 12
- Pages
- 6742-6746
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W1974248650
- Cited by
- 564
- References
- 1
- FWCI
- 5.10
- Type
- journal article
- Language
- en
- Open access
- green
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- Ellis, H. M., Yu, D., DiTizio, T., & Court, D. L. (2001). High efficiency mutagenesis, repair, and engineering of chromosomal DNA using single-stranded oligonucleotides. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.121164898
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