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A phage integrase directs efficient site-specific integration in human cells
Open accessAbstract
The integrase from the Streptomyces phage φC31 carries out efficient recombination between the attP site in the phage genome and the attB site in the host bacterial chromosome. In this paper, we show that the enzyme also functions in human cells. A plasmid assay system was constructed that measured intramolecular integration of attP into attB . This assay was used to demonstrate that in the presence of the φC31 integrase, precise unidirectional integration occurs with an efficiency of 100% in Escherichia coli and >50% in human cells. This assay system was also used to define the minimal sizes of attB and attP at 34 bp and 39 bp, respectively. Furthermore, precise and efficient intermolecular integration of an incoming plasmid bearing attP into an established Epstein–Barr virus plasmid bearing attB was documented in human cells. This work is a demonstration of efficient, site-specific, unidirectional integration in mammalian cells. These observations form the basis for site-specific integration strategies potentially useful in a broad range of genetic engineering applications.
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Cited by
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Siuti, P., Yazbek, J., & Lu, T. K. (2013) · Nature Biotechnology · doi:10.1038/nbt.2510 · Cited by 531
Phage as agents of lateral gene transfer
Canchaya, C., Fournous, G., Chibani-Chennoufi, S., Dillmann, M.-L., & Brüssow, H. (2003) · Current Opinion in Microbiology · doi:10.1016/s1369-5274(03)00086-9 · Cited by 499
Chen, Z.-Y., He, C.-Y., Ehrhardt, A., & Kay, M. A. (2003) · Molecular Therapy · doi:10.1016/s1525-0016(03)00168-0 · Cited by 462
Site-specific integrase-mediated transgenesis in mice via pronuclear injection
Tasic, B., Hippenmeyer, S., Wang, C., Gamboa, M., Zong, H., Chen-Tsai, Y., & Luo, L. (2011) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1019507108 · Cited by 246
Metadata
- Authors
- Groth, A. C., Olivares, E. C., Thyagarajan, B., & Calos, M. P.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 2000-05-09
- Volume
- 97
- Issue
- 11
- Pages
- 5995-6000
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W1988379532
- Cited by
- 495
- References
- 0
- FWCI
- 4.62
- Type
- journal article
- Language
- en
- Open access
- green
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- Groth, A. C., Olivares, E. C., Thyagarajan, B., & Calos, M. P. (2000). A phage integrase directs efficient site-specific integration in human cells. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.090527097
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