Intracellular delivery of nucleic acids using pressure

US5922687A · kind A · utility

Assignee

• Board of Trustees of the Leland Stanford Junior University · US

Inventors

• Michael J. Mann · San Francisco, US
• Frank P. Diet · Neunburg vorm Wald, DE
• Victor J. Dzau · Newton, US
• Gary H. Gibbons · Atlanta, US
• Heiko Von Der Leyen · Sehnde, DE

Key dates

• Filing date: Nov 7, 1996
• Grant date: Jul 13, 1999
• Expiry date: Nov 7, 2016

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N2320/32
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Naked nucleic acids (DNA, RNA, and/or analogs), drugs, and/or other molecules in an extracellular environment enter cells in living intact tissue upon application of pressure to the cells and extracellular environment. Nucleic acids localize to the cell nuclei. Transfection efficiencies greater than 90% are achievable for naked DNA and RNA. A sealed enclosure, defined by an enclosing means and/or tissue, contains the cells and their extracellular environment. The enclosure is pressurized to an incubation pressure on the order of atmospheres. A protective inelastic sheath may be used to prevent distension and trauma in tissue that is part of the enclosure boundary. Suitable enclosures include pressurization chambers and organs such as blood vessels or the heart. Parts of organs, entire organs, and/or entire organisms are pressurized. Suitable target tissue types include blood vessel (in particular vein) tissue, heart, kidney, liver, and bone marrow tissue. Gene therapy applications include ex-vivo treatment of grafts prior to transplantation, and in-vivo treatment of tissue. Useful therapy targets include cell cycle regulatory genes for blocking cell proliferation, and interleukin (…