Defective sindbis viral vectors
Disclosed herein are new defective Sindbis viral vectors made from a novel Helper plasmid, with differences in envelope proteins between JT vectors and consensus Sindbis virus sequences, and also between JT and Ar-339 vectors. Also disclosed are vectors produced using the plasmid, methods for producing the vectors, methods for treating mammals suffering from tumors and pharmaceutical formulations for use in the treatment methods.
1. An isolated nucleic acid comprising a nucleotide sequence consisting of the sequence as set forth in SEQ ID NO: 60 (T7-DM H101).
2. An isolated nucleic acid comprising a nucleotide sequence consisting of the sequence as set forth in SEQ ID NO: 65 T7-Stul-RLacZ#202.
3. A method for producing defective Sindbis virus vectors comprising the steps of
(a) providing a linearized Replicon plasmid comprising a nucleotide sequence consisting of the sequence as set forth in SEQ ID NO: 65 and a linearized Helper plasmid comprising a nucleotide sequence consisting of the sequence as set forth in SEQ ID NO: 60;
(b) transcribing said Replicon plasmid and said Helper plasmid to produce RNA;
(c) collecting the RNA transcribed in step (b) from said Replicon plasmid and said Helper plasmid and transfecting cells with said RNA;
(d) incubating said transfected cells for a time and at a temperature effective for producing defective Sindbis viral vectors; and
(e) collecting said defective Sindbis viral vectors from the medium of said transfected cells.