Epigenetic regulation of gene structure and function with a cell-permeable Cre recombinase.
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.
Jo
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Department of Microbiology and Immunology, Vanderbilt University School of Medicine, 1161 21 Avenue South, AA4210, Nashville, TN 37232-2363, USA.
Nashabi
A
A
Doxsee
C
C
Lin
Q
Q
Unutmaz
D
D
Chen
J
J
Ruley
H E
HE
D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D051519:Fibroblast Growth Factor 4; D005346:Fibroblast Growth Factors / Q000235:genetics / Q000378:metabolism; D019426:Integrases / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014161:Transduction, Genetic; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism
C496446:Fgf4 protein, mouse; D051519:Fibroblast Growth Factor 4; D011518:Proto-Oncogene Proteins; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D005346:Fibroblast Growth Factors; C045073:Cre recombinase; D019426:Integrases