Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-9333227-B2
Publication Date
2016-05-10
Expiration Date
Abstract
It can be useful to regulate the growth of microbial cells. Some embodiments herein provide genetically engineered microbial cells that can produce bacteriocins to control the growth of microbial cells. In some embodiments, microbial cells are contained within a desired environment. In some embodiments, contaminating microbial cells are neutralized. In some embodiments, a first microbial cell type regulates the growth of a second microbial cell type so as to maintain a desired ratio of the two cell types.
Core Innovation
The invention relates to a genetically engineered microbial cell that controls growth of a second microbial cell in a culture medium by secreting a bacteriocin. The secreted bacteriocin inhibits or prevents reproduction of the second microbial cell, and it is capable of inhibiting or preventing reproduction of the genetically engineered microbial cell in the absence of a cell immunity modulator that confers resistance to the secreted bacteriocin.
The genetically engineered microbial cell includes a first gene expression system that expresses the secreted bacteriocin encoded by a first nucleic acid sequence under the control of a first promoter. The cell also includes a second gene expression system comprising an expression vector with a second nucleic acid sequence that encodes the cell immunity modulator, which confers resistance to the secreted bacteriocin to protect the genetically engineered microbial cell.
The second gene expression system is genetically engineered to permit transcription, post-transcriptional expression, and post-translational activity of the cell immunity modulator when neutralizing an undesired second microbial cell is desired, while decreasing or eliminating at least one of transcription, post-transcriptional expression, or post-translational activity when immunity is not desired. The second gene expression system includes at least one of a second promoter inactive concurrent with expression of the secreted bacteriocin, a nucleic acid encoding a transcriptional repressor, a ribozyme or antisense oligonucleotide complementary to at least a portion of the second nucleic acid sequence, or a regulatable tRNA specific for a transcript of the second nucleic acid sequence.
Claims Coverage
The independent claim set defines a bacteriocin-based growth-control system with 4 inventive features, including a secreted bacteriocin expression system, a plasmid-based immunity modulator expression vector, multi-level immunity control, and conditional neutralization in the culture medium.
Secreted bacteriocin controlled growth inhibition of a second microbial cell
A genetically engineered microbial cell having a first nucleic acid sequence under a first promoter that encodes a secreted bacteriocin capable of inhibiting or preventing reproduction of the genetically engineered microbial cell in the absence of a cell immunity modulator that confers resistance to the secreted bacteriocin.
Immunity-modulator expression vector with conditional transcription, post-transcriptional expression, and post-translational activity
A second gene expression system comprising an expression vector on a plasmid that encodes a cell immunity modulator, wherein the cell immunity modulator confers resistance to the secreted bacteriocin and the second gene expression system permits transcription, post-transcriptional expression, and post-translational activity of the cell immunity modulator.
Immunity modulation reduction during bacteriocin expression
The second gene expression system is capable of decreasing or eliminating at least one of transcription of the immunity-modulator nucleic acid, post-transcriptional expression of the cell immunity modulator, or post-translational activity of the cell immunity modulator, using a second promoter inactive concurrent with bacteriocin expression, a transcriptional repressor, a ribozyme or antisense oligonucleotide, or a regulatable tRNA.
Neutralization condition enabling bacteriocin inhibition in the culture medium
When neutralizing an undesired second microbial cell is desired, the first gene expression system is active and the second gene expression system permits transcription, post-transcriptional expression, and post-translational activity of the cell immunity modulator, thereby allowing the secreted bacteriocin to inhibit or prevent reproduction of the second microbial cell in the culture medium.
Across the independent claim, the system combines a secreted bacteriocin with a cell immunity modulator whose transcription and/or post-transcriptional and/or post-translational activity is engineered to be decreased or eliminated during bacteriocin expression, with promoter inactivity, transcriptional repression, ribozyme or antisense silencing, or regulatable tRNA control.
Stated Advantages
Allows the secreted bacteriocin to inhibit or prevent reproduction of a second microbial cell in a culture medium.
Provides resistance to the secreted bacteriocin for the genetically engineered microbial cell via a cell immunity modulator.
Enables decreasing or eliminating cell immunity modulator transcription, post-transcriptional expression, or post-translational activity, including through engineered promoter inactivity, transcriptional repressor, ribozyme or antisense oligonucleotide, or regulatable tRNA.
Supports conditional operation so that, when neutralizing an undesired second microbial cell is desired, bacteriocin inhibition occurs while immunity modulator activity is permitted.
Documented Applications
Controlling growth of a second microbial cell in a culture medium by secreting a bacteriocin from a genetically engineered microbial cell.
Neutralizing an undesired second microbial cell in the culture medium by activating bacteriocin gene expression while permitting immunity modulator activity.
Interested in licensing this patent?