Abstract 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. Intracellular delivery of cargo molecules according to this invention is accomplished by the use of novel transport polypeptides which comprise HIV tat protein or one or more portions thereof, and which are covalently attached to cargo molecules. The transport polypeptides in preferred embodiments 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. By virtue of the absence of the cysteine-rich region, the preferred transport polypeptides of this invention solve the potential problems of spurious trans-activation and disulfide aggregation. The reduced size of the preferred transport polypeptides of this invention also minimizes interference with the biological activity of the cargo molecule.
A fusion protein comprising a carboxy-terminal cargo moiety and an amino-terminal transport moiety, wherein the cargo moiety consists of amino acids 43-412 of HSV VP16 protein and the transport moiety consists of amino acids 47-58 of HIV tat protein.
Claim 2
Dependent Claim
39 words
A fusion protein comprising a carboxy-terminal cargo moiety and an amino-terminal transport moiety, wherein the cargo moiety consists of amino acids 43-412 of HSV VP16 protein and the transport moiety consists of amino acids 47-58 of HIV tat protein.
Claim 3
Dependent Claim
17 words
The fusion protein according to claim 1, wherein the transport moiety is preceded by an amino-terminal methionine.
Claim 4
Dependent Claim
14 words
A composition comprising the fusion protein of claim 1 or 2 and a carrier.
Claim 5
Dependent Claim
6 words
Fusion protein JB106 (SEQ ID NO:38).
Claim 6
Dependent Claim
6 words
Fusion protein JB106 (SEQ ID NO:38).
Claim 7
Dependent Claim
6 words
Fusion protein JB117 (SEQ ID NO:59).
Claim 8
Dependent Claim
6 words
Fusion protein JB117 (SEQ ID NO:59).
Claim 9
Dependent Claim
6 words
Fusion protein JB118 (SEQ ID NO:60).
Claim 10
Dependent Claim
6 words
Fusion protein JB118 (SEQ ID NO:60).
Claim 11
Dependent Claim
6 words
Fusion protein JB119 (SEQ ID NO:61).
Claim 12
Dependent Claim
6 words
Fusion protein JB119 (SEQ ID NO:61).
Claim 13
Dependent Claim
6 words
Fusion protein JB120 (SEQ ID NO:62).
Claim 14
Dependent Claim
6 words
Fusion protein JB120 (SEQ ID NO:62).
Claim 15
Dependent Claim
6 words
Fusion protein JB122 (SEQ ID NO:63).
Claim 16
Dependent Claim
6 words
Fusion protein JB122 (SEQ ID NO:63).
Claim 17
Dependent Claim
18 words
A composition comprising the fusion protein of claim 4, 5, 6, 7, 8 or 9 and a carrier.