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Proc Natl Acad Sci U S A. Jan 18, 1994; 91(2): 664–668.
PMCID: PMC43009

Tat-mediated delivery of heterologous proteins into cells.

Abstract

The Tat protein of human immunodeficiency virus 1 (HIV-1) can enter cells efficiently when added exogenously in tissue culture. To assess if Tat can carry other molecules into cells, we chemically cross-linked Tat peptides (residues 1-72 or 37-72) to beta-galactosidase, horseradish peroxidase, RNase A, and domain III of Pseudomonas exotoxin A (PE) and monitored uptake colorimetrically or by cytotoxicity. The Tat chimeras were effective on all cell types tested, with staining showing uptake into all cells in each experiment. In mice, treatment with Tat-beta-galactosidase chimeras resulted in delivery to several tissues, with high levels in heart, liver, and spleen, low-to-moderate levels in lung and skeletal muscle, and little or no activity in kidney and brain. The primary target within these tissues was the cells surrounding the blood vessels, suggesting endothelial cells, Kupffer cells, and/or splenic macrophages. Tat-mediated uptake may allow the therapeutic delivery of macromolecules previously thought to be impermeable to living cells.

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  • Foldvari M, Mezei C, Mezei M. Intracellular delivery of drugs by liposomes containing P0 glycoprotein from peripheral nerve myelin into human M21 melanoma cells. J Pharm Sci. 1991 Nov;80(11):1020–1028. [PubMed]
  • Chakrabarti R, Wylie DE, Schuster SM. Transfer of monoclonal antibodies into mammalian cells by electroporation. J Biol Chem. 1989 Sep 15;264(26):15494–15500. [PubMed]
  • Connor J, Huang L. Efficient cytoplasmic delivery of a fluorescent dye by pH-sensitive immunoliposomes. J Cell Biol. 1985 Aug;101(2):582–589. [PMC free article] [PubMed]
  • Ortiz D, Baldwin MM, Lucas JJ. Transient correction of genetic defects in cultured animal cells by introduction of functional proteins. Mol Cell Biol. 1987 Aug;7(8):3012–3017. [PMC free article] [PubMed]
  • McNeil PL, Murphy RF, Lanni F, Taylor DL. A method for incorporating macromolecules into adherent cells. J Cell Biol. 1984 Apr;98(4):1556–1564. [PMC free article] [PubMed]
  • Renneisen K, Leserman L, Matthes E, Schröder HC, Müller WE. Inhibition of expression of human immunodeficiency virus-1 in vitro by antibody-targeted liposomes containing antisense RNA to the env region. J Biol Chem. 1990 Sep 25;265(27):16337–16342. [PubMed]
  • Wu GY, Wu CH. Delivery systems for gene therapy. Biotherapy. 1991;3(1):87–95. [PubMed]
  • Prior TI, FitzGerald DJ, Pastan I. Translocation mediated by domain II of Pseudomonas exotoxin A: transport of barnase into the cytosol. Biochemistry. 1992 Apr 14;31(14):3555–3559. [PubMed]
  • Prior TI, FitzGerald DJ, Pastan I. Barnase toxin: a new chimeric toxin composed of pseudomonas exotoxin A and barnase. Cell. 1991 Mar 8;64(5):1017–1023. [PubMed]
  • Stenmark H, Moskaug JO, Madshus IH, Sandvig K, Olsnes S. Peptides fused to the amino-terminal end of diphtheria toxin are translocated to the cytosol. J Cell Biol. 1991 Jun;113(5):1025–1032. [PMC free article] [PubMed]
  • Ishihara H, Hara T, Aramaki Y, Tsuchiya S, Hosoi K. Preparation of asialofetuin-labeled liposomes with encapsulated human interferon-gamma and their uptake by isolated rat hepatocytes. Pharm Res. 1990 May;7(5):542–546. [PubMed]
  • Basu SK. Receptor-mediated endocytosis of macromolecular conjugates in selective drug delivery. Biochem Pharmacol. 1990 Nov 1;40(9):1941–1946. [PubMed]
  • Wu GY, Wu CH. Evidence for targeted gene delivery to Hep G2 hepatoma cells in vitro. Biochemistry. 1988 Feb 9;27(3):887–892. [PubMed]
  • Wilson JM, Grossman M, Wu CH, Chowdhury NR, Wu GY, Chowdhury JR. Hepatocyte-directed gene transfer in vivo leads to transient improvement of hypercholesterolemia in low density lipoprotein receptor-deficient rabbits. J Biol Chem. 1992 Jan 15;267(2):963–967. [PubMed]
  • Khorana HG. Rhodopsin, photoreceptor of the rod cell. An emerging pattern for structure and function. J Biol Chem. 1992 Jan 5;267(1):1–4. [PubMed]
  • Leamon CP, Low PS. Delivery of macromolecules into living cells: a method that exploits folate receptor endocytosis. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5572–5576. [PMC free article] [PubMed]
  • Wu GY, Wilson JM, Shalaby F, Grossman M, Shafritz DA, Wu CH. Receptor-mediated gene delivery in vivo. Partial correction of genetic analbuminemia in Nagase rats. J Biol Chem. 1991 Aug 5;266(22):14338–14342. [PubMed]
  • Kumagai AK, Eisenberg JB, Pardridge WM. Absorptive-mediated endocytosis of cationized albumin and a beta-endorphin-cationized albumin chimeric peptide by isolated brain capillaries. Model system of blood-brain barrier transport. J Biol Chem. 1987 Nov 5;262(31):15214–15219. [PubMed]
  • Frankel AD, Pabo CO. Cellular uptake of the tat protein from human immunodeficiency virus. Cell. 1988 Dec 23;55(6):1189–1193. [PubMed]
  • Mann DA, Frankel AD. Endocytosis and targeting of exogenous HIV-1 Tat protein. EMBO J. 1991 Jul;10(7):1733–1739. [PMC free article] [PubMed]
  • Studier FW, Rosenberg AH, Dunn JJ, Dubendorff JW. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. [PubMed]
  • Melby EL, Prydz K, Olsnes S, Sandvig K. Effect of monensin on ricin and fluid phase transport in polarized MDCK cells. J Cell Biochem. 1991 Nov;47(3):251–260. [PubMed]
  • Dang CV, Lee WM. Nuclear and nucleolar targeting sequences of c-erb-A, c-myb, N-myc, p53, HSP70, and HIV tat proteins. J Biol Chem. 1989 Oct 25;264(30):18019–18023. [PubMed]
  • Calnan BJ, Biancalana S, Hudson D, Frankel AD. Analysis of arginine-rich peptides from the HIV Tat protein reveals unusual features of RNA-protein recognition. Genes Dev. 1991 Feb;5(2):201–210. [PubMed]
  • Lord JM, Hartley MR, Roberts LM. Ribosome inactivating proteins of plants. Semin Cell Biol. 1991 Feb;2(1):15–22. [PubMed]
  • Stirpe F, Wawrzynczak EJ, Brown AN, Knyba RE, Watson GJ, Barbieri L, Thorpe PE. Selective cytotoxic activity of immunotoxins composed of a monoclonal anti-Thy 1.1 antibody and the ribosome-inactivating proteins bryodin and momordin. Br J Cancer. 1988 Nov;58(5):558–561. [PMC free article] [PubMed]
  • FitzGerald D, Pastan I. Redirecting Pseudomonas exotoxin. Semin Cell Biol. 1991 Feb;2(1):31–37. [PubMed]
  • Hwang J, Fitzgerald DJ, Adhya S, Pastan I. Functional domains of Pseudomonas exotoxin identified by deletion analysis of the gene expressed in E. coli. Cell. 1987 Jan 16;48(1):129–136. [PubMed]
  • Ensoli B, Barillari G, Salahuddin SZ, Gallo RC, Wong-Staal F. Tat protein of HIV-1 stimulates growth of cells derived from Kaposi's sarcoma lesions of AIDS patients. Nature. 1990 May 3;345(6270):84–86. [PubMed]
  • Vogel J, Hinrichs SH, Reynolds RK, Luciw PA, Jay G. The HIV tat gene induces dermal lesions resembling Kaposi's sarcoma in transgenic mice. Nature. 1988 Oct 13;335(6191):606–611. [PubMed]
  • Jaye M, Howk R, Burgess W, Ricca GA, Chiu IM, Ravera MW, O'Brien SJ, Modi WS, Maciag T, Drohan WN. Human endothelial cell growth factor: cloning, nucleotide sequence, and chromosome localization. Science. 1986 Aug 1;233(4763):541–545. [PubMed]
  • Auron PE, Webb AC, Rosenwasser LJ, Mucci SF, Rich A, Wolff SM, Dinarello CA. Nucleotide sequence of human monocyte interleukin 1 precursor cDNA. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7907–7911. [PMC free article] [PubMed]
  • Acland P, Dixon M, Peters G, Dickson C. Subcellular fate of the int-2 oncoprotein is determined by choice of initiation codon. Nature. 1990 Feb 15;343(6259):662–665. [PubMed]
  • Abraham JA, Mergia A, Whang JL, Tumolo A, Friedman J, Hjerrild KA, Gospodarowicz D, Fiddes JC. Nucleotide sequence of a bovine clone encoding the angiogenic protein, basic fibroblast growth factor. Science. 1986 Aug 1;233(4763):545–548. [PubMed]
  • Weeks BS, Desai K, Loewenstein PM, Klotman ME, Klotman PE, Green M, Kleinman HK. Identification of a novel cell attachment domain in the HIV-1 Tat protein and its 90-kDa cell surface binding protein. J Biol Chem. 1993 Mar 5;268(7):5279–5284. [PubMed]
  • Vogel BE, Lee SJ, Hildebrand A, Craig W, Pierschbacher MD, Wong-Staal F, Ruoslahti E. A novel integrin specificity exemplified by binding of the alpha v beta 5 integrin to the basic domain of the HIV Tat protein and vitronectin. J Cell Biol. 1993 Apr;121(2):461–468. [PMC free article] [PubMed]

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