Eureka AIR delivers breakthrough ideas for toughest innovation challenges, trusted by R&D personnel around the world.

Synthetic chimeric vaccinia virus

A vaccinia virus, virus technology, applied in the direction of viruses, antiviral agents, virus antigen components, etc., can solve the problems of no scrape-free area status regulations and guidelines, prion contamination, etc.

Pending Publication Date: 2021-03-23
戴维·埃文斯 +2
View PDF6 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is a risk of contamination with bovine spongiform encephalopathy or scrapie prions
Although regulations and guidelines for modern vaccines state that all materials used must come from BSE-free areas, there are no regulations and guidelines regarding the status of scrapie-free areas.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Synthetic chimeric vaccinia virus
  • Synthetic chimeric vaccinia virus
  • Synthetic chimeric vaccinia virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0162] Example 1. Selection and Design of Overlapping Fragments of Viral Genomes

[0163] Synthetic chimeric VACV ACAM2000 containing VACV WR strain hairpin and duplex sequences (scVACV ACAM2000-WR DUP / HP)

[0164] The design of the scVACV genome was based on the previously described genome sequence of VACV ACAM2000 [GenBank accession number AY313847] (Osborne JD et al. Vaccine. 2007;25(52):8807-32). The genome was divided into nine overlapping segments (Fig. 1). These fragments were designed such that they shared at least 1.0 kbp of overlapping sequence (ie, homology) with each adjacent fragment to provide sites where homologous recombination would drive full-length genome assembly (Table 1). These overlapping sequences provide sufficient homology for accurate recombination between co-transfected fragments (Yao XD, Evans DH. Journal of Virology. 2003;77(13):7281-90).

[0165] Table 1: VACV ACAM2000 genomic fragments used in this study. The size and sequence within the VACV...

Embodiment 2

[0176] Ligation of VACV WR F and S-terminal hairpin loops to VACV ACAM2000 right and left ITR fragments

[0177] Synthesize the same 70bp repeat fragment as VACV WR strain ( Figure 2C ; SEQ ID NO: 10). SapI and NheI restriction sites were included at the 5' and 3' ends of the 70 bp tandem repeat to facilitate ligation to the VACV WR hairpin sequence and the VACVACAM2000 right and left ITR fragments, respectively. Before the VACV WR terminal hairpin loop can be ligated onto the 70bp tandem repeat, the loop must be extended by an additional 58bp using a duplex sequence synthesized by IDT technology ( Figure 3A ). This is due to the extra sequence immediately downstream of the concatemer resolution site, before the first 70 bp repeat found in VACV strain WR. The duplex sequence is produced by synthesizing two single-stranded DNA molecules that, when annealed together, will produce a 5'-TGT overhang at the 5' end and a 5'-GGT overhang at the 3' end end of the duplex DNA ( ...

Embodiment 3

[0182] Example 3. Preparation of VACV ACAM2000 overlapping DNA fragments

[0183] Each VACV ACAM2000 overlapping DNA fragment in Table 1 was cloned into the plasmid provided by GeneArt using the restriction enzyme I-SceI. Before transfecting these synthetic DNA fragments into BGMK cells, the plasmid was digested with I-SceI and the product was run on a gel to confirm that the DNA fragments were successfully linearized ( Figure 5). After digestion at 37°C for 2 hours, the reaction was subsequently heat inactivated at 65°C. Samples were kept on ice or at 4°C until formation of terminal hairpins / duplexes / 70bp tandem repeats / ITR fragments (as described above).

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

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 syntheticchimeric vaccinia viruses. The synthetic chimeric vaccinia viruses are well suited, among others, as virus vaccines or to generate an oncolytic response and pharmaceutical formulations.

Description

Background technique [0001] The Sequence Listing associated with this application is filed electronically in text format via EFS-Web and is hereby incorporated by reference in its entirety into the specification. The name of the text file containing the sequence listing is 104545-0031-WO-SequenceListing.txt. The text file was created on May 2, 2019 and is 288,652 bytes in size. [0002] Poxviruses (members of the Poxviridae family) are double-stranded DNA viruses that can infect both humans and animals. Poxviruses are divided into two subfamilies based on host range. The subfamily Chordopoxviridae, which infects vertebrate hosts, consists of eight genera, four of which are known (Orthopoxvirus, Parapoxvirus, Molluscumpoxvirus ( Molluscipoxvirus) and Yatapoxvirus) infect humans. Smallpox is caused by infection with variola virus (VARV), a member of the genus Orthopoxvirus (OPV). The OPV genus includes a number of genetically related and morphologically identical viruses, i...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01A61K39/275
CPCC12N7/00A61K39/285C12N2710/24143C12N2710/24121A61P31/20A61K2039/525A61P35/00C12N2710/24151C12R2001/91C12N1/00A61K39/275C12N2710/24134A61K39/12C12N2710/24122C12N2710/24132A61K35/768A61K2039/5252A61K2039/80
Inventor 戴维·埃文斯瑞安·诺伊斯塞思·莱德曼
Owner 戴维·埃文斯
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Eureka Blog
Learn More
PatSnap group products