Non-replicative transduction particles and transduction particle-based reporter systems
AssigneeGeneWeave Biosciences, Inc.
InventorDiego Ariel Rey
Life of this patent
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Abstract
Methods and systems are provided for packaging reporter nucleic acid molecules into non-replicative transduction particles for use as reporter molecules. The non-replicative transduction particles can be constructed from viruses and use viral transduction and replication systems. The reporter nucleic acid molecules include a reporter gene, such as a reporter molecule or selectable marker, for detecting target genes or cells. Methods and systems are provided for detection of cells and target nucleic acid molecules using the non-replicative transduction particles as reporter molecules.
Claims14
A bacterial cell packaging system for packaging a reporter nucleic acid molecule into a non-replicative transduction particle (NRTP) for introduction into a cell, the packaging system comprising:
- a host bacteria cell;
- a first nucleic acid construct inside the host bacteria cell, comprising of a bacteriophage genome having a non-functional packaging initiation site sequence, wherein the non-functional packaging initiation site sequence prevents packaging of the bacteriophage genome into the NRTP;
- and
- a second nucleic acid construct inside the host bacteria cell and separate from the first nucleic acid construct, comprising of the reporter nucleic acid molecule having a reporter gene and a functional packaging initiation site sequence for facilitating packaging of a replicon of the reporter nucleic acid molecule into the NRTP,
- wherein the functional packaging initiation site sequence on the second nucleic acid construct complements the non-functional packaging initiation site sequence in the bacteriophage genome on the first nucleic acid construct.
A bacterial cell packaging system for packaging a reporter plasmid into a non-replicative transduction particle (NRTP) for introduction into a cell, the packaging system comprising:
- a host bacteria cell;
- a first nucleic acid construct inside the host bacteria cell, comprising of a φ80α bacteriophage genome having a non-functional φ80α small terminase (terS) gene, wherein the non-functional φ80α terS gene prevents packaging of the φ80α bacteriophage genome into the NRTP;
- and
- a second nucleic acid construct inside the host bacteria cell and separate from the first nucleic acid construct, comprising of the reporter plasmid having a luxAB reporter gene operatively linked to a S. aureus clpB promoter comprising the sequence of SEQ ID NO:
- 12, a pT181 origin of replication comprising the sequence of SEQ ID NO:
- 5, and a functional φ80α terS gene comprising the sequence of SEQ ID NO:
- 9, the functional φ80α terS gene facilitating packaging of a replicon of the reporter nucleic acid molecule into the NRTP,
- wherein the functional φ80α terS gene on the second nucleic acid construct complements the non-functional φ80α terS gene in the φ80α bacteriophage genome on the first nucleic acid construct.
Dependencies are inferred from the claim text.