Tat-mediated delivery of heterologous proteins into cells.
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.
Fawell
S
S
Biogen Inc., Cambridge, MA 02142.
Seery
J
J
Daikh
Y
Y
Moore
C
C
Chen
L L
LL
Pepinsky
B
B
Barsoum
J
J
D036002:ADP Ribose Transferases; D000595:Amino Acid Sequence; D000818:Animals; D001427:Bacterial Toxins; D001693:Biological Transport, Active; D002352:Carrier Proteins / Q000235:genetics / Q000493:pharmacokinetics*; D005098:Exotoxins / Q000235:genetics / Q000493:pharmacokinetics; D015696:Gene Products, tat / Q000235:genetics / Q000493:pharmacokinetics*; D015497:HIV-1 / Q000378:metabolism*; D006367:HeLa Cells; D006735:Horseradish Peroxidase / Q000493:pharmacokinetics; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics / Q000493:pharmacokinetics; D011506:Proteins / Q000008:administration & dosage / Q000493:pharmacokinetics* / Q000627:therapeutic use; D012259:Ribonuclease, Pancreatic / Q000493:pharmacokinetics; D014018:Tissue Distribution; D037521:Virulence Factors; D001616:beta-Galactosidase / Q000493:pharmacokinetics; D054322:tat Gene Products, Human Immunodeficiency Virus; D000097668:Pseudomonas aeruginosa Exotoxin A
D036002:ADP Ribose Transferases; D001427:Bacterial Toxins; D002352:Carrier Proteins; D005098:Exotoxins; D015696:Gene Products, tat; D006735:Horseradish Peroxidase; D010446:Peptide Fragments; D011506:Proteins; D012259:Ribonuclease, Pancreatic; D037521:Virulence Factors; D001616:beta-Galactosidase; D054322:tat Gene Products, Human Immunodeficiency Virus; D000097668:Pseudomonas aeruginosa Exotoxin A