US 7,320,859 B2
Modular transfection systems
Hanns-Martin Schmidt, Köln (Germany); Ludger Altrogge, Pulheim (Germany); Dietmar Lenz, Köln (Germany); Gudula Riemen, Langenfeld (Germany); Helmut Brosterhus, Kirchundern (Germany); Elke Lorbach, Köln (Germany); Juliana Helfrich, Köln (Germany); Katharina Hein, Köln (Germany); Marion Gremse, Köln (Germany); Tatjana Males, Hilden (Germany); Rainer Christine, Köln (Germany); Gregor Siebenkotten, Frechan-Königsdorf (Germany); Bodo Ortmann, Köln (Germany); Tamara Turbanski, Wiehl (Germany); and Andreas Klaes, Köln (Germany)
Assigned to Amaxa AG, (Germany)
Appl. No. 10/466,368
PCT Filed Jan. 10, 2002, PCT No. PCT/DE02/00060
§ 371(c)(1), (2), (4) Date Aug. 13, 2003,
PCT Pub. No. WO02/055721, PCT Pub. Date Jul. 18, 2002.
Claims priority of application No. 101 00 996 (DE), filed on Jan. 10, 2001.
Prior Publication US 2004/0137622 A1, Jul. 15, 2004
Int. Cl. C12Q 1/68 (2006.01)
U.S. Cl. 435—6  [435/455; 435/194; 530/350] 15 Claims
 
1. A method for transfection of cells using at least one protein capable of forming nucleoprotein filaments, wherein the protein is initially modified with at least two functional components of different function, and/or wherein different proteins are each initially modified with components of different function, each functional component influencing one or more steps of the transfection, a nucleic acid to be transfected is then loaded with the modified protein, the nucleic acid and the protein forming a filament-like complex, and this complex is added to the cells to be transfected, wherein
a. the protein binds a nucleic acid sequence and forms a nucleoprotein filament, and
b. the protein so modified is modified by the addition of any of a functional component which causes association of the complex to the cellular surface, endosomal or non-endosomal passage of the complex through the cell membrane, release of the complex from endosomes or lysosomes, transport of the complex into the nucleus, or combination thereof,
thereby facilitating transfection.