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Recombination System Based on Cre α Complementation and Leucine Zipper Fusions

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Abstract

In a previous study, a recombination system based on the α complementation of cre recombinase and protein transduction was established. This system relied on the transient expression of the inactive, self-excisable C-terminal (β) and the transduction of the N-terminal (α) cre fragments to cells as a purified protein. This recombination system potentially results in a less invasive and more controllable cre recombination in mammalian cells. In this study, we have employed a more efficient complementation triggering sequence using more than only the overlapping amino acids to help the α and β fragments reassociate. In order to increase the association efficiency of the complementing fragments of cre recombinase, we chose to use a fusion of cre fragments to a self-heterodimerizing pair of proteins to trigger their binding and thus increase the efficiency of the restored enzymatic activity. For this purpose, the leucine zipper motifs (bJun and bFos) of the AP-1 transcription were fused to cre fragments (α and β, respectively). This resulted in an increased reassociation efficiency of the fragments and a two times more efficient recombination system compared with the previous study.

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References

  1. Sauer, B., & Henderson, N. (1988). Proceedings of the National Academy of Sciences of the United States of America, 85, 5166–5170.

    Article  CAS  Google Scholar 

  2. Nagy, A. (2000). Genesis, 26, 99–109.

    Article  CAS  Google Scholar 

  3. Breckenridge, R., Kotecha, S., Towers, N., Bennett, M., & Mohun, T. (2007). Genesis, 45, 135–144.

    Article  CAS  Google Scholar 

  4. Schmidt, E. E., Taylor, D. S., Prigge, J. R., Barnett, S., & Capecchi, M. R. (2000). Proceedings of the National Academy of Sciences of the United States of America, 97, 13702–13707. doi:10.1073/pnas.240471297.

    Article  CAS  Google Scholar 

  5. Loonstra, A., Vooijs, M., Beverloo, H. B., Allak, B. A., van Drunen, E., Kanaar, R., et al. (2001). Proceedings of the National Academy of Sciences of the United States of America, 98, 9209–9214. doi:10.1073/pnas.161269798.

    Article  CAS  Google Scholar 

  6. Silver, D. P., & Livingston, D. M. (2001). Molecular Cell, 8, 233–243. doi:10.1016/S1097-2765(01)00295-7.

    Article  CAS  Google Scholar 

  7. Casanova, E., Lemberger, T., Fehsenfeld, S., Mantamadiotis, T., & Schutz, G. (2003). Genesis, 37, 25–29.

    Article  CAS  Google Scholar 

  8. Jullien, N., Sampieri, F., Enjalbert, A., & Herman, J. P. (2003). Nucleic Acids Research, 31, e131. doi:10.1093/nar/gng131.

    Article  Google Scholar 

  9. Seidi, A., Mie, M., & Kobatake, E. (2007). Biotechnology Letters, 29, 1315–1322. doi:10.1007/s10529-007-9406-6.

    Article  CAS  Google Scholar 

  10. Glover, J. N., & Harrison, S. C. (1995). Nature, 373, 257–261. doi:10.1038/373257a0.

    Article  CAS  Google Scholar 

  11. Hess, J., Angel, P., & Pchorpp-Kistner, M. (2004). Journal of Cell Science, 117, 5965–5973. doi:10.1242/jcs.01589.

    Article  CAS  Google Scholar 

  12. Smeal, T., Angel, P., Meek, J., & Karin, M. (1989). Genes & Development, 3, 2091–2100. doi:10.1101/gad.3.12b.2091.

    Article  CAS  Google Scholar 

  13. Xu, Y., Xu, G., Liu, B., & Gu, G. (2007). Nucleic Acids Research, 35, e126. doi:10.1093/nar/gkm559.

    Article  Google Scholar 

  14. Ghosh, I., Hamilton, A. D., & Regan, L. (2000). Journal of the American Chemical Society, 122, 5658–5659. doi:10.1021/ja994421w.

    Article  CAS  Google Scholar 

  15. Hu, C. D., Chinencov, Y., & Kerppola, T. (2002). Molecular Cell, 9, 789–798. doi:10.1016/S1097-2765(02)00496-3.

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama, 226-8501, Japan

    Azadeh Seidi, Masayasu Mie & Eiry Kobatake

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  1. Azadeh Seidi
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  2. Masayasu Mie
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  3. Eiry Kobatake
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Correspondence to Eiry Kobatake.

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Seidi, A., Mie, M. & Kobatake, E. Recombination System Based on Cre α Complementation and Leucine Zipper Fusions. Appl Biochem Biotechnol 158, 334–342 (2009). https://doi.org/10.1007/s12010-008-8409-7

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  • DOI: https://doi.org/10.1007/s12010-008-8409-7

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