Propagator cells and methods for propagating phage, in particular for delivering CRISPR-Cas components via probiotic organisms
AssigneeSNIPR Technologies Limited
InventorJasper Clube
Life of this patent
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Abstract
The invention provides propagator cells and methods for propagating phage and transduction particles.
Claims21
A method of producing a population of phages, wherein the phages are of a first type capable of infecting host cells of a first bacterial species or strain by binding a cell-surface receptor comprised by bacteria of the first species or strain, the method comprising:
- (a) providing a population of second bacterial cells comprising the receptor on the surface of the second cells, wherein the second cells are of a second species or strain, wherein the second species or strain is different from the first species or strain;
- (b) infecting the second cells with the phages of the first type;
- and
- (c) propagating the phages in the second cells, thereby producing the population of phages,
- wherein the phages comprise a nucleotide sequence encoding crRNAs that are operable with a Cas in bacteria of the host cell strain or species to form an active CRISPR/Cas system that is capable of targeting one or more protospacer nucleotide sequences, wherein each target sequence is comprised by the genome of the host cells, whereby the crRNAs guide the Cas in host cells to modify the target sequence, thereby killing host cells or reducing host cell population growth;
- and
- wherein the genome of each second bacterial cell does not comprise the target sequence.
A method of producing a population of phages, wherein the phages are of a first type capable of infecting host cells of a first bacterial species or strain by binding a cell-surface receptor comprised by bacteria of the first species or strain, the method comprising
- (a) providing a population of second bacterial cells comprising the receptor on the surface of the second cells, wherein the second cells are of a second species or strain, wherein the second species or strain is different from the first species or strain;
- (b) infecting the second cells with the phages of the first type;
- and
- (c) propagating the phages in the second cells, thereby producing the population of phages;
- wherein the phages comprise a nucleotide sequence encoding crRNAs that are operable with a Cas in bacteria of the host cell strain or species to form an active CRISPR/Cas system that is capable of targeting one or more protospacer nucleotide sequences, wherein each target sequence is comprised by the genome of the host cells, whereby the crRNAs guide the Cas in host cells to modify the target sequence, thereby killing host cells or reducing host cell population growth;
- and
- wherein when infected by the phage, the second cells do not comprise the active CRISPR/Cas system.
A method of producing a population of phages, wherein the phages are of a first type capable of infecting host cells of a first bacterial species or strain by binding a cell-surface receptor comprised by bacteria of the first species or strain, the method comprising
- (a) providing a population of second bacterial cells comprising the receptor on the surface of the second cells, wherein the second cells are of a second species or strain, wherein the second species or strain is different from the first species or strain;
- (b) infecting the second cells with the phages of the first type;
- and
- (c) propagating the phages in the second cells, thereby producing the population of phages;
- wherein the phages comprise a nucleotide sequence encoding crRNAs that are operable with a Cas in bacteria of the host cell strain or species to form an active CRISPR/Cas system that is capable of targeting one or more protospacer nucleotide sequences, wherein each target sequence is comprised by the genome of the host cells, whereby the crRNAs guide the Cas in host cells to modify the target sequence, thereby killing host cells or reducing host cell population growth;
- and
- wherein the crRNAs comprise repeat sequences that are not operable with a Cas of the second cells.
A method of producing a population of phages, wherein the phages are of a first type capable of infecting host cells of a first bacterial species or strain by binding a cell-surface receptor comprised by bacteria of the first species or strain, the method comprising
- (a) providing a population of second bacterial cells comprising the receptor on the surface of the second cells, wherein the second cells are of a second species or strain, wherein the second species or strain is different from the first species or strain;
- (b) infecting the second cells with the phages of the first type;
- and
- (c) propagating the phages in the second cells, thereby producing the population of phages;
- wherein the second bacterial cells are engineered to produce the receptor, wherein wild-type bacteria of the second species or strain do not produce the receptor.
A method of producing a population of phages, wherein the phages are of a first type capable of infecting host cells of a first bacterial species or strain by binding a cell-surface receptor comprised by bacteria of the first species or strain, the method comprising
- (a) providing a population of second bacterial cells comprising the receptor on the surface of the second cells, wherein the second cells are of a second species or strain, wherein the second species or strain is different from the first species or strain;
- (b) infecting the second cells with phages of the first type;
- and
- (c) propagating the phages in the second cells, thereby producing the population of phages;
- and
- wherein the receptor comprises a teichoic acid moiety that is the product of the action of one or more enzymes in the second cell, wherein the genome of the second cell comprises one or more expressible nucleotide sequences encoding the one or more enzymes, wherein wild-type cells of the second species or strain do not comprise the expressible nucleotide sequences.
Dependencies are inferred from the claim text.