Lentiviral triplex dna, and vectors and recombinant cells...

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/867 (2006.01) A61K 48/00 (2006.01) C12N 5/10 (2006.01) C12N 7/01 (2006.01)

Patent

CA 2382832

The present ivention provides nucleic acid, vectors, viruses, and recombinant cells comprising triple-stranded structures, such as those resulting from central initiation and termination of HIV-1 reverse transcription at the center of HIV-1 linear DNA genomes. These triplex structures can act as a cis- determinant of HIV-1 DNA nuclear import, allowing infection of non-dividing target cells. In one aspect, the presence of the DNA triplex sequence in an HIV vector strongly stimulates gene transfer in hematopoietic stem cells. The invention also provides methods of using these triplex structures for making recombinant cells, as well as methods of using the recombinant cells to express proteins of interest both in vitro and in vivo.

L'invention concerne un acide nucléique, des vecteurs, des virus, et des cellules de recombinaison comprenant des structures à triple brin, telles que celles résultant d'une initiation centrale et d'une termination de transcription inverse du VIH-1 au niveau du centre des génomes d'ADN linéaire du VIH-1. Ces structures triplex peuvent agir comme un cis-déterminant d'importation nucléaire d'ADN du VIH-1, ce qui entraîne l'infection de cellules cibles qui ne se divisent pas. Selon un aspect de l'invention, la présence de la séquence d'ADN triplex dans un vecteur du VIH stimule fortement un transfert génique dans des cellules souches hématopoïétiques. Cette invention concerne également des techniques d'utilisation de ces structures triplex pour produire des cellules de recombinaison, ainsi que des techniques utilisant lesdites cellules de recombinaison pour exprimer des protéines considérées à la fois in vitro et in vivo.

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