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Epigenetic regulation of gene structure and function with a cell-permeable Cre recombinase

Nature Biotechnology volume 19, pages 929–933 (2001)Cite this article

Abstract

Studies of mammalian gene function are hampered by temporal limitations in which phenotypes occurring at one stage of development interfere with analysis at later stages. Moreover, phenotypes resulting from altered gene activity include both direct and indirect effects that may be difficult to distinguish. In the present study, recombinant fusion proteins bearing the 12 amino acid membrane translocation sequence (MTS) from the Kaposi fibroblast growth factor (FGF-4) were used to transduce enzymatically active Cre proteins directly into mammalian cells. High levels of recombination were observed in a variety of cultured cell types and in all tissues examined in mice following intraperitoneal administration. This represents the first use of protein transduction to induce the enzymatic conversion of a substrate in living cells and animals and provides a rapid and efficient means to manipulate mammalian gene structure and function.

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Figure 1: Recombinant Cre Proteins and intracellular uptake.
Figure 2: In vivo Cre-mediated recombination.
Figure 3: Cre-mediated recombination ex vivo.
Figure 4: Cre-mediated recombination in mice.
Figure 5: Histological examination of β-galactosidase expression.
Figure 6: Time course of lacZ expression in splenocytes and thymocytes from Cre-treated mice.

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Acknowledgements

We thank Mark Magnuson and Yao Zhong Lin for providing S4R ES cells and pMTS2 plasmid, respectively, and Jacek Hawiger, Dean Ballard, and Gene Oltz for critical reading of this manuscript. This work was supported by a Public Health Service Grant (R01RR13166 to H.E.R.) and by a grant from the Kleberg Foundation. Additional support was provided by a Cancer Center Support grant (P30CA42014) to the Vanderbilt-Ingram Cancer Center.

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  1. Department of Microbiology and Immunology, Vanderbilt University School of Medicine, 1161 21 Avenue South, AA4210, Nashville, 37232-2363, TN

    Daewoong Jo, Abudi Nashabi, Christie Doxsee, Derya Unutmaz & H. Earl Ruley

  2. Department of Medicine, Vanderbilt University School of Medicine, 1161 21 Avenue South, AA4210, Nashville, 37232-2363, TN

    Qing Lin & Jin Chen

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  1. Daewoong Jo
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Correspondence to H. Earl Ruley.

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Jo, D., Nashabi, A., Doxsee, C. et al. Epigenetic regulation of gene structure and function with a cell-permeable Cre recombinase. Nat Biotechnol 19, 929–933 (2001). https://doi.org/10.1038/nbt1001-929

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