Tat-derived transport polypeptides

US6316003A · kind A · utility

Assignees

• Whitehead Institute For Biomedical Research · US
• Johns Hopkins Univ. School of Medicine · US
• Biogen Inc. · US

Inventors

• Alan Frankel · Tiburon, US
• Carl O. Pabo · Newton, US
• James Barsoum · Lexington, US
• Stephen E. Fawell · Winchester, US
• R. Blake Pepinsky · Arlington, US

Key dates

• Filing date: Apr 28, 1994
• Grant date: Nov 13, 2001
• Expiry date: Apr 28, 2014

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N2740/16322
• WIPO field: Pharmaceuticals
• WIPO sector: Chemistry

Abstract

This invention relates to delivery of biologically active cargo molecules, such as polypeptides and nucleic acids, into the cytoplasm and nuclei of cells in vitro and in vivo. Intracellular delivery of cargo molecules according to this invention is accomplished by the use of novel transport polypeptides which include HIV tat protein or one or more portions thereof, and which are covalently attached to cargo molecules. The transport polypeptides in preferred embodiments of this invention are characterized by the presence of the tat basic region (amino acids 49-57), the absence of the tat cysteine-rich region (amino acids 22-36) and the absence of the tat exon 2-encoded carboxy-terminal domain (amino acids 73-86) of the naturally-occurring tat protein. By virtue of the absence of the cysteine-rich region, the preferred transport polypeptides of this invention solve the potential problems of spurious trans-activation and disulfide aggregation. The reduced size of the preferred transport polypeptides of this invention also minimizes interference with the biological activity of the cargo molecule.