Novel vesicular stomatitis virus and virus rescue system

Patent metadata

Field	Value
Publication number	US-2020172932-A1
Application number	US-201916717360-A
Country	US
Kind code	A1
Filing date	Dec 17, 2019
Priority date	Sep 21, 2011
Publication date	Jun 4, 2020
Grant date	—

Abstract

The present relation relates to recombinant vesicular stomatitis virus for use as prophylactic and therapeutic vaccines as well as the preparation and purification of immunogenic compositions which are formulated into the vaccines of the present invention.

First claim

What is claimed is:

1 . A vesicular stomatitis virus (VSV) genomic clone comprising:
(a) a VSV genome encoding and expressing a nucleocapsid, phosphoprotein, matrix, glycoprotein and large protein, wherein the VSV genome comprises nucleotide substitutions and amino acid coding changes to improve replicative fitness and genetic stability, (b) a cloning vector, (c) an extended T7 promoter, (d) a hammerhead ribozyme, (e) a hepatitis delta virus ribozyme and T7 terminator (f) unique restriction endonuclease cleavage sites in a VSV genomic sequence (g) a leader and a trailer that are cis-acting sequences controlling mRNA synthesis and replication

2 . The VSV genomic clone of claim 1 , wherein the cloning vector is pSP72 (Genbank X65332.2)

3 . The VSV genomic clone of claim 1 , wherein the extended T7 promoter is PT7-g10.

4 . The VSV genomic clone of claim 1 , wherein the unique restriction endonuclease cleavage sites are 1367 NheI, 2194 SpeI, 2194 BstBI, 4687 PacI, 7532 AvaI, 10190 SalI and 11164 AflII.

5 . The VSV genomic clone of claim 1 , wherein the VSV genomic clone is depicted in FIG. 1 .

6 . The VSV genomic clone of claim 1 , wherein the nucleotide position is according to GenBank Accession Number EF197793 and wherein the nucleotide substitutions are selected from the group consisting of
1371 CA>GC (NheI) After 2195 insert TAG (SpeI) (all genome numbers below adjusted to include +3 bp) 3036 G>T improves match to consensus transcription stop signal 3853 X>A (was an ambiguity in Genbank file) 4691 T>A to generate Pad 7546 C>A silent change in L coding sequence eliminates a BstBI site 1960 TAC>TCC to change Y>S 3247 GTA>ATA to change V>I 3729 AAG>GAG to change K>E 4191 GTA>GAA to change V>E 4386 GGT>GAT to change G>D 4491 ACC>ATC to change T>I 5339 ATT>CTT to change I>L 5834 ACT>GCT to change T>A and 10959 AGA>AAA to change R>K.

7 . The VSV genomic clone of claim 1 , wherein the nucleotide position is according to GenBank Accession Number EF197793 and wherein the nucleotide substitutions are selected from the group consisting of:

Nucleotide position in
Nucleotide position
Nucleotide

EF197793
in rEF197793
Change
Purpose

1
Substitution 1371-2
Substitution 1371-2
CA > GC
Creates a unique NheI cleavage site

between N and P gens

2
Substitution 1960-2
Substitution 1960-2
TAC > TCC
Y > S substitution in P protein amino

acid sequence to agree with

consensus.

3
Insert after 2195
3 base insert after
Insert TAG
Creates a unique SpeI site between P

2195

and M genes

4
Substitution 3039
Substitution 3042
G > T
Improves agreement with consensus.

Also improves agreement with

consensus transcription stop signal

5
Substitution 3234-6
Substitution 3237-9
GTA > ATA
V > I substitution in P protein amino

acid sequence to agree with consensus.

6
Substitution 3729-31
Substitution 3732-34
AAG > GAG
K > E substitution in G protein amino

acid sequence to agree with

consensus.

7
Substitution 3856
Substitution 3859
N > A
Replace unknown base in Genbank

file with consensus

8
Substitution 4191-93
Substitution 4194-6
GTA > GAA
V > E substitution in G protein amino

acid sequence to agree with

consensus.

9
Substitution 4386-88
Substitution 4389-92
GGT > GAT
G > D substitution in G protein amino

acid sequence to agree with

consensus.

10
Substitution 4491-93
Substitution 4494-96
ACC > ATC
T > I substitution in G protein amino

acid sequence to agree with consensus.

11
Substitution 4694
Substitution 4697
T > A
Creates unique PacI cleavage site

between G and L genes

12
Substitution 5339-41
Substitution 5342-44
ATT > CTT
I > L substitution in L protein amino

acid sequence to agree with consensus.

Assignees

• Int Aids Vaccine Initiative

Inventors

• Parks Christopher L
• Yuan Maoli
• Wright Kevin
• Jurgens Christy

Classifications

• C12N2800/22

  Vectors comprising a coding region that has been codon optimised for expression in a respective host · CPC title

• C12N2760/20243

  viral genome or elements thereof as genetic vector · CPC title

• C12N7/00

  Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) · CPC title

• C12N15/86 Primary

  Viral vectors · CPC title

• C12N2760/20221

  Viruses as such, e.g. new isolates, mutants or their genomic sequences · CPC title