Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Recombinant nucleic acid molecules, expression cassettes and bacteria, and methods of use thereof

A technology for recombining nucleic acids and expression cassettes, applied in chemical instruments and methods, bacteria, bacterial peptides, etc., can solve the problem of unsuccessful attenuation or inactivation of pathogens or cells, and the attenuation of sensors or cancer cells cannot be guaranteed with certainty, etc. question

Inactive Publication Date: 2012-02-08
ADURO BIOTECH
View PDF21 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Heterologous antigen delivery is particularly advantageous for the treatment or prevention of diseases or conditions caused, inter alia, by toxic or lethal sources such as cancer or pathogens (e.g., HIV or hepatitis B), where injection of natural sensors or cancer cells affects the affected organisms are potentially harmful and administration of attenuated or inactivated pathogens or cells has proven unsuccessful in eliciting an effective immune response, or where adequate attenuation of the sensor or cancer cells cannot be assured with certainty

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
  • Recombinant nucleic acid molecules, expression cassettes and bacteria, and methods of use thereof
  • Recombinant nucleic acid molecules, expression cassettes and bacteria, and methods of use thereof
  • Recombinant nucleic acid molecules, expression cassettes and bacteria, and methods of use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 28

[0282] Alternatively, other known intergenic regions or synthetic sequences can be used to construct polycistronic expression cassettes for use in Listeria or other bacteria. Construction of intergenic regions leading to substantial secondary RNA structure should be prevented to avoid unwanted termination of transcription by rho-independent mechanisms.

[0283] Importantly, signal peptides must functionally link each coding region if secretion of any or all translated proteins expressed from polycistronic messages is desired. In some embodiments, these signal peptides are different from each other.

[0284] Thus, in some embodiments, the expression cassettes described herein for use in Listeria or other bacteria are polycistronic (eg, dicistronic). Two or more polypeptides are encoded as discrete polypeptides by a bicistronic or polycistronic expression cassette. In some embodiments, the bicistronic or polycistronic expression cassette includes an intergenic sequence (eg, fr...

Embodiment 1

[0453] Example 1. Preparation of Exemplary Mutant Listeria Strains

[0454] The Listeria strain was derived from 10403S (Bishop et al., J. Immunol. 139:2005 (1987)). Listeria strains with in-frame deletions of the indicated genes were generated by SOE-PCR and allelic exchange using well-established methods (Camilli, et al., Mol. Microbiol. 8:143 (1993)). Mutant LLO L461T (DP-L4017) is described in Glomski et al., J. Cell. Biol. 156:1029 (2002), incorporated herein by reference. actA - The mutant strain (DP-L4029) is the DP-L3078 strain described in Skoble et al., J. of Cell Biology, 150:527-537 (2000), which is hereby incorporated by reference in its entirety, and whose prophage has self-healed. (Prophage self-healing is described in (Lauer et al., J. Bacteriol. 184:4177 (2002); US Patent Publication No. 2003 / 0203472)). actA - wxya - The construction of strains is described in US Provisional Application 60 / 446,051, filed February 6, 2003, as L4029 / uvrAB (see, eg, Exampl...

Embodiment 2

[0456] Example 2. Construction of Listeria strains expressing AH1 / OVA or AH1-A5 / OVA

[0457] A mutant Listeria strain expressing a truncated form of the model antigen ovalbumin (OVA), an immunodominant epitope called AH1 (SPSYVYHQF (SEQ ID NO :72)); and altered epitopes AH1-A5 (SPSYAYHQF (SEQ ID NO:73); Slansky et al., Immunity, 13:529-538 (2000)). The pPL2 integrating vector (Lauer et al., J Bacteriol. 184:4177 (2002); U.S. Patent Publication No. 2003 / 0203472) was used to generate a single copy of OVA and AH1- A5 / OVA recombinant Listeria strain.

[0458] A. Construction of OVA-expressing Listeria (DP-L4056)

[0459] An antigen expression cassette consisting of hemolysin-deleted LLO fused to truncated OVA and contained in the pPL2 integration vector (pPL2 / LLO-OVA) was first made. By attaching tRNA at the PSA (phage from ScottA) Arg -attBB' Introduction of pPL2 / LLO-OVA into phage self-healing Listeria monocytogenes strain DP-L4056 to generate Listeria-OVA vaccine strain. ...

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 present invention provides recombinant nucleic acid molecules, expression cassettes and vectors for expressing polypeptides including heterologous polypeptides such as antigens in bacteria. Some recombinant nucleic acid molecules, expression cassettes and vectors include codon-optimized sequences encoding polypeptides and / or signal peptides. Some recombinant nucleic acid molecules, expression cassettes and expression vectors include sequences encoding non-Listeria and / or non-secA1 signal peptides for secreted polypeptides. The present invention also provides bacteria comprising nucleic acid molecules, expression cassettes and expression vectors, and compositions such as vaccines comprising the bacteria. Methods of making and using the bacteria, recombinant nucleic acid molecules and expression cassettes are also provided.

Description

[0001] Cross References to Related Applications [0002] Pursuant to 35 U.S. C. § 119(e) this application claims the benefit of priority to each of the following U.S. provisional applications, the disclosures of each of which are hereby incorporated by reference in their entirety: U.S. Provisional Application Serial No. 60 / 616,705, titled "Bacterial Expression Cassette, Bacterial Vaccine Composition and Method of Use thereof", Thomas W. Dubensky, Jr et al., filed October 6, 2004 (Docket No. 282173003923); U.S. Provisional Application Serial No. 60 / 615,287, Title of Invention "Bacterial Expression Cassette, Bacterial Vaccine Composition and Method of Use thereof", Thomas W.Dubensky, Jr, etc., filed on October 1, 2004 (Docket No.282173003922); U.S. Provisional Application Serial No.60 / 599,377, 2004 Filed August 5; U.S. Provisional Application Serial No. 60 / 556,744, filed March 26, 2004; U.S. Provisional Application Serial No. 60 / 541,515, filed February 2, 2004; and U.S. Provisiona...

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 Patents(China)
IPC IPC(8): A61K39/00A61K39/02A61P31/00A61K48/00C07H21/04C07K14/195C12N1/21C12N15/74
CPCC07K2319/02A61K2039/523A61K39/00C07K14/195
Inventor T·W·小杜本斯基D·A·波特努瓦W·S·小勒基特D·N·库克
Owner ADURO BIOTECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com