Tat-derived transport polypeptides

Details Tat-derived transport polypeptides Tat-derived transport polypeptides

C - Chemistry, Metallurgy

12

N

C12N 15/62 (2006.01) A61K 39/385 (2006.01) A61K 47/48 (2006.01) C07K 14/025 (2006.01) C07K 14/16 (2006.01) C07K 14/21 (2006.01) C07K 14/47 (2006.01) C07K 19/00 (2006.01) C12N 9/08 (2006.01) C12N 9/22 (2006.01) C12N 15/49 (2006.01) C12N 15/87 (2006.01) A61K 38/00 (2006.01) A61K 39/00 (2006.01)

Patent

CA 2135642

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 by the use of novel transport polypeptides which comprise one or more portions of HIV tat protein and which are covalently attached to cargo molecules. The transport polypeptides 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. The absence of the cysteine-rich region found in conventional tat proteins solves the problems of spurious trans-activation and disulfide aggregation.

La présente invention concerne l'apport de molécules cargo biologiquement actives, telles que des polypeptides et des acides nucléiques dans le cytoplasme et les noyaux de cellules in vitro et in vivo, grâce à l'emploi de nouveaux polypeptides de transport qui comportent une ou plusieurs parties de la protéine tat du VIH et qui se fixent par covalence aux molécules cargo. Ces polypeptides de transport sont caractérisés par la présence de la région basique tat (acides aminés 49-57), l'absence de la région riche en cystéine (acides aminés 22-36) et l'absence du domaine carboxy-terminal, encodé en position 2 par l'exon tat (acides aminés 73-86), de la protéine tat se rencontrant à l'état naturel. L'absence de la région riche en cystéine constatée dans les protéines tat classiques résout les problèmes de fausse transactivation et d'agrégation des ponts disulfure.

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