Host defense suppressing methods and compositions for modulating a genome
AssigneeFlagship Pioneering Innovations VI, LLC
InventorsRobert James CITORIK, Jacob Rosenblum RUBENS, Cecilia Giovanna Silvia COTTA-RAMUSINO, William Edward SALOMON, Zi Jun WANG
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Abstract
The disclosure provides, e.g., compositions and methods for modulating a host response to a Gene Writer system. In some embodiments, modulation of the host response results in increased integration of a heterologous nucleic acid sequence of interest into a target genome. In some embodiments, modulation of the host response results in an increased stability, e.g., maintenance of an insertion or expression thereof. In some embodiments, modulation of the host response results in decreased cytotoxicity.
Claims32
A method of modifying a target DNA molecule in a mammalian host cell, the method comprising:
- a) contacting the host cell with a gene modifying system;
- and
- b) contacting the host cell with a host response modulator,
- wherein the gene modifying system comprises a Gene Writer polypeptide, or a nucleic acid encoding the Gene Writer polypeptide, and a template nucleic acid, the template nucleic acid comprising i) a sequence that binds the Gene Writer polypeptide and ii) a heterologous object sequence;
- wherein the Gene Writer polypeptide comprises a Cas9 nickase and the RT domain from a retrovirus.
A kit comprising:
- a) a gene modifying system that comprises a Gene Writer polypeptide, or a nucleic acid encoding the Gene Writer polypeptide, and a template nucleic acid, the template nucleic acid comprising i) a sequence that binds the Gene Writer polypeptide and ii) a heterologous object sequence;
- and
- b) a host response modulator;
- wherein the Gene Writer polypeptide comprises a Cas9 nickase and the RT domain from a retrovirus.
A composition comprising:
- a) a gene modifying system that comprises a Gene Writer polypeptide, or a nucleic acid encoding the Gene Writer polypeptide, and a template nucleic acid, the template nucleic acid comprising i) a sequence that binds the Gene Writer polypeptide and ii) a heterologous object sequence;
- and
- b) a host response modulator;
- wherein the Gene Writer polypeptide comprises a Cas9 nickase and the RT domain from a retrovirus.
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Dependencies are inferred from the claim text.