Transbiotic Regulation of Bacterial Gene Expression
AssigneePebble Labs USA Inc.
InventorsRichard Sayre, Tatiana Vinogradova-Shah, Elena Sineva, Pedro Costa-Nunes
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Abstract
The inventive technology may include novel systems for regulation of the expression of bacterial genes through the introduction of antisense RNA (asRNA) that may disrupt expression of targeted pathogenic genes and/or their products (mRNA, proteins). In some embodiments, the inventive technology may include novel genetically engineered donor bacterial strains that are configured to efficiently and continuously deliver asRNA polynucleotides to a recipient pathogen and downregulate expression or one or more essential genes.
Claims33
1-41. (canceled)
A method of controlling gene expression in pathogenic bacteria comprising the steps of:
- generating a genetically modified donor bacteria configured to express a heterologous asRNA polynucleotide directed to an essential gene of a bacterial pathogen;
- introducing said genetically modified donor bacteria to a target host that is infected with said bacterial pathogen, or is susceptible to infection by said bacterial pathogen;
- expressing said heterologous asRNA polynucleotide directed to an essential gene of said bacterial pathogen;
- transporting said heterologous asRNA polynucleotide directed to an essential gene of said bacterial pathogen out of said genetically modified donor bacteria;
- introducing said heterologous asRNA polynucleotide directed to an essential gene of said bacterial pathogen wherein said bacterial pathogen takes up said asRNA polynucleotide;
- and
- inhibiting expression of said essential gene of bacterial pathogen through the action of said heterologous asRNA polynucleotide hybridizing with the mRNA of said essential gene of a bacterial pathogen.
A method of controlling bacterial biofilm formation comprising the steps:
- generating a genetically modified donor bacteria configured to express a heterologous asRNA polynucleotide directed to an essential gene that contributes to biofilm formation by a bacterial pathogen;
- introducing said genetically modified donor bacteria to a target host that is infected with said bacterial pathogen, or is susceptible to infection by said bacterial pathogen;
- expressing said heterologous asRNA polynucleotide directed to an essential gene that contributes to biofilm formation by said bacterial pathogen;
- transporting said heterologous asRNA polynucleotide directed to an essential gene that contributes to biofilm formation by said bacterial pathogen out of said genetically modified donor bacteria;
- introducing said heterologous asRNA polynucleotide directed to an essential gene that contributes to biofilm formation by said bacterial pathogen wherein said bacterial pathogen takes up said asRNA polynucleotide;
- and
- inhibiting expression of said essential gene that contributes to biofilm formation by a bacterial pathogen through the action of said heterologous asRNA polynucleotide hybridizing with the complementary mRNA of said essential gene of a bacterial pathogen.
A method of treating a Vibrio infection in an organism comprising the steps of:
- generating a genetically modified donor bacteria configured to express a heterologous asRNA polynucleotide that is complementary to the mRNA of DNA adenine methylase (Dam) of a Vibrio bacterial pathogen;
- introducing said genetically modified donor bacteria to a target host that is infected with said Vibrio bacterial pathogen, or is susceptible to infection by said Vibrio bacterial pathogen;
- expressing said heterologous asRNA polynucleotide that is complementary to said mRNA of DNA adenine methylase (Dam) of said Vibrio bacterial pathogen;
- transporting said heterologous asRNA polynucleotide that is complementary to the mRNA of DNA adenine methylase (Dam) of said Vibrio bacterial pathogen out of said genetically modified donor bacteria;
- introducing said heterologous asRNA polynucleotide that is complementary to the mRNA of DNA adenine methylase (Dam) of said Vibrio bacterial pathogen wherein said bacterial pathogen takes up said asRNA polynucleotide;
- and
- inhibiting expression of said essential gene of bacterial pathogen through the action of said asRNA hybridizing with the complementary mRNA of said essential gene of said Vibrio bacterial pathogen.
Dependencies are inferred from the claim text.