Improved methods of transfection

Details Improved methods of transfection Improved methods of transfection

C - Chemistry, Metallurgy

12

N

C12N 15/88 (2006.01) C07K 7/06 (2006.01) C07K 7/08 (2006.01) C07K 16/28 (2006.01) C12N 5/10 (2006.01) C12N 15/87 (2006.01) C12P 21/00 (2006.01)

Patent

CA 2410419

Transfection of confluent cells or other slowly dividing or non-dividing cells that are in contact with each other with a nucleic acid using a non-viral receptor targeted vector may be improved by the concurrent use of an agent that disrupts cell-cell junctions, especially EGTA. The vector is especially an integrin-targeting transfection vector complex comprising (i) a nucleic acid, especially a nucleic acid encoding a sequence of interest, (ii) an integrin-binding component, especially an integrin-targeting peptide, (iii) a polycationic nucleic acid-binding component, especially an oligolysine, and (iv) a lipid component, especially, DOPE, DOTMA, DOSPA or combinations thereof. Various applications of the improved method of transfection are described.

L'invention concerne la transfection de cellules confluentes ou autres cellules à division lente ou sans division, en contact mutuel, par le biais d'un acide nucléique dépourvu de vecteur ciblé sur récepteur non viral, sachant que cette transfection peut être améliorée par l'utilisation concurrente d'un agent qui défait les liens entre cellules, en particulier l'EGTA. En particulier, le vecteur est un complexe à vecteur de transfection ciblant l'intégrine qui comprend (i) un acide nucléique, notamment un acide nucléique codant une séquence visée, (ii) un composant de liaison avec l'intégrine, notamment un peptide ciblant l'intégrine, (iii) un composant de liaison avec un acide nucléique polycationique, notamment une oligosine, et (iv) un composant lipide, notamment DOPE, DOTMA, DOSPA ou des combinaisons de ces éléments. L'invention concerne par ailleurs diverses applications du procédé de transfection amélioré qui est décrit.

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