In vivo protein transduction: Delivery of a biologically active protein into the mouse

Science

Volumn 285, Issue 5433, 1999, Pages 1569-1572

  Schwarze, Steven R ^a   Ho, Alan ^a   Vocero Akbani, Adamina ^a   Dowdy, Steven F ^a  

^a WASHINGTON UNIVERSITY SCHOOL OF MEDICINE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

BETA GALACTOSIDASE; HYBRID PROTEIN; TRANSACTIVATOR PROTEIN;

ANIMAL EXPERIMENT; ARTICLE; BLOOD BRAIN BARRIER; BRAIN TISSUE; CONTROLLED STUDY; DRUG BRAIN LEVEL; DRUG DELIVERY SYSTEM; DRUG PENETRATION; DRUG UPTAKE; ENZYME ACTIVITY; GENETIC COMPLEMENTATION; GENETIC TRANSDUCTION; HUMAN IMMUNODEFICIENCY VIRUS; INTRAPERITONEAL DRUG ADMINISTRATION; MOUSE; NONHUMAN; PRIORITY JOURNAL;

ANIMALS; BETA-GALACTOSIDASE; BLOOD-BRAIN BARRIER; BRAIN; CELL MEMBRANE; DRUG CARRIERS; DRUG DELIVERY SYSTEMS; FLUORESCEIN-5-ISOTHIOCYANATE; GENE PRODUCTS, TAT; HUMANS; INJECTIONS, INTRAPERITONEAL; JURKAT CELLS; LIPID BILAYERS; MICE; MICE, INBRED C57BL; MICROSCOPY, CONFOCAL; MICROSCOPY, FLUORESCENCE; MUSCLE, SKELETAL; RECOMBINANT FUSION PROTEINS; SPLEEN; TISSUE DISTRIBUTION; TUMOR CELLS, CULTURED;

ANIMALIA; HUMAN IMMUNODEFICIENCY VIRUS;

EID: 0033520487     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.285.5433.1569     Document Type: Article

References (32)

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14. Sixty-five 4-to 8-week-old C57BL/6 mice were injected intraperitoneally with 1.7 nmol of TAT-FITC peptide or with control FITC in 500 μl of phosphate-buffered saline (PBS) or 100 to 500 μg of TAT-β-Gal or β-Gal control protein in 0.5 to 2.0 ml of PBS and 10% glycerol. Whole blood cells were isolated from the orbital artery, and splenocytes were isolated at indicated time points and analyzed by FACS. Treated mice were killed, and tissues were harvested and frozen in Histo Prep media (Fisher Scientific). Sections (10 to 50 μm) were cut on a cryostat, fixed in 0.25% gluteraldehyde for 15 min, and developed for 4 hours (liver, kidney, lung, heart, and spleen) or 16 hours (brain) in 0.2% X-Gal solution (10) or analyzed by fluorescence confocal microscopy. All mice injected with either TAT-FITC peptide or TAT-β-Gal yielded positive results. All animal procedures were performed in accordance with institutional guidelines.
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32. We thank C. Turck (Univ. of California, San Francisco) for peptide synthesis, T. Woodford-Thomas for confocal microscopy, L. White for animal care, and T. Woolsey and all the members of the Dowdy lab for critical input. S.R.S. was supported by an NIH training grant, and A.H. was supported by an NIH-Medical Scientist Training Program training grant. This work was supported by the Howard Hughes Medical Institute.