Synthetic chimeric vaccinia virus
AssigneeTonix Pharma Limited
InventorsDavid Evans, Ryan Noyce, Seth Lederman
Life of this patent
Estimating the term…
Abstract
The invention relates in various aspects to a synthetic chimeric vaccinia virus or compositions comprising such viruses, and the development and use of systems and methods for producing such synthetic chimeric vaccinia viruses. The synthetic chimeric vaccinia viruses are well suited, among others, as virus vaccines or to generate an oncolytic response and pharmaceutical formulations.
Claims15
A synthetic chimeric vaccinia virus (scVACV) that is replicated and reactivated from DNA derived from chemically synthesized DNA, the genome of said scVACV comprising one or more modifications to eliminate one or more unique restriction sites or to add or repair one or more unique restriction sites relative to a wild type vaccinia virus genome, wherein the scVACV further comprises a left and a right terminal hairpin loop derived from chemically synthesized DNA, one or both of the terminal hairpin loops being heterologous relative to the wild type vaccinia virus genome, and wherein the one or both of the terminal hairpin loops comprise different tandem repeat regions or a different number thereof relative to the wild type form of the one or both of the terminal hairpin loops.
A method of producing a synthetic chimeric vaccinia virus (scVACV) that is replicated and reactivated from DNA derived from chemically synthesized DNA comprising the steps of:
- (i) chemically synthesizing DNA fragments, the DNA fragments corresponding to the DNA that is replicated and reactivated and comprising one or more modifications to eliminate one or more unique restriction sites or to add or repair one or more unique restriction sites relative to a wild type vaccinia virus genome;
- (ii) chemically synthesizing DNA corresponding to a left and a right terminal hairpin loop, one or both loops being heterologous relative to the wild type vaccinia virus genome and wherein one or both of the terminal hairpin loops comprise different tandem repeats regions, or a different number thereof, than a corresponding wild type terminal hairpin loop, and ligating the terminal hairpin loops derived from the chemically synthesized terminal hairpin loops to the corresponding left termini and right termini of the DNA fragments derived from the chemically synthesized DNA of step (i);
- (iii) transfecting the ligated DNA fragments of step (ii) into helper virus-infected cells;
- (iv) culturing said cells to produce a mixture of helper virus and scVACV particles in said cells;
- and
- (v) plating the mixture on host cells specific to the scVACV to recover the scVACV.
Dependencies are inferred from the claim text.