Targeting pseudotyped retroviral vectors
The present invention provides targeted lentiviral vectors that are psuedotyped with mutated Sindbis envelopes. For example, mutations in the Sindbis E2 protein are used to alter viral titer, specificity, specificity index, tropism, and susceptibility to host immune response. Typically, one or more of the E1, E2, or E3 proteins can be mutated at one or more amino acid positions. The psuedotyped, targeted lentiviral vectors of the invention are used to transduce heterologous genes into a cell and can be used for in vivo and ex vivo therapeutic applications, as well as for diagnostic and research tool applications.
1. An isolated pseudotyped retrovirus comprising mutant Sindbis virus E2 and E3 proteins, wherein the sequence SLKQ of the Sindbis virus E2 protein is substituted or deleted and wherein the sequence RSKR of the Sindbis virus E3 protein is substituted or deleted.
2. The pseudotyped retrovirus of claim 1 , further comprising a targeting moiety.
3. The pseudotyped retrovirus claim 2 , wherein the targeting moiety is an antibody directed against a tumor specific cell surface antigen.
4. The pseudotyped retrovirus of claim 3 , wherein the targeting moiety is an antibody directed against p-glycoprotein.
5. The pseudotyped retrovirus of claim 3 , wherein the targeting moiety is an antibody directed against the transferrin receptor.
6. The isolated pseudotyped retrovirus of claim 1 further comprising a mutant Sindbis virus E1 protein wherein the mutant Sindbis virus E1 is the E1 protein encoded by SEQ ID NO:5.
7. The isolated pseudotyped retrovirus of claim 1 wherein the mutant E2 protein is the protein encoded by SEQ ID NO:3.
8. The isolated pseudotyped retrovirus of claim 1 wherein the sequence SLKQ of the Sindbis virus E2 protein is substituted with the amino acid sequence AAAA.
9. The isolated pseudotyped retrovirus of claim 1 wherein the sequence SLKQ of the Sindbis virus E2 protein is deleted.
10. The isolated pseudotyped retrovirus of claim 1 wherein the sequence RSKR of the Sindbis virus E3 protein is substituted with the amino acid sequence AAAA.
11. The isolated pseudotyped retrovirus of claim 1 wherein the sequence RSKR of the Sindbis virus E3 protein is deleted.
12. The isolated pseudotyped retrovirus of claim 1 wherein the sequence SLKQ of the Sindbis virus E2 protein is substituted and the sequence RSKR of the Sindbis virus E3 protein is deleted.
13. An isolated pseudotyped retrovirus comprising mutant Sindbis virus protein E2 and E3, wherein the sequence SLKQ of the Sindbis virus E2 protein is substituted with the amino acid sequence AAAA and wherein the sequence RSKR of the Sindbis virus E3 protein is deleted.