DNA compositions and related methods
AssigneeFlagship Pioneering Innovations Vii, LLC
InventorsAlexandra Rachael Sneider, Jacob Rosenblum Rubens, Narahari Subbanna Pujar, Molly Krisann Gibson, Benjamin Andrew Portney, Geoffrey A. Von Maltzahn, Camilo Ayala Breton
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Abstract
The disclosure provides, for example, single stranded, covalently closed DNA that does not form a double stranded structure longer than 100 base pairs. The ssDNA may encode an effector sequence, for instance a therapeutic protein. The ssDNA may comprise a nuclear targeting sequence (NTS). In some embodiments, the ssDNA shows decreased activation of the innate immune system compared to an otherwise similar dsDNA.
Claims22
A pharmaceutical formulation comprising:
- a lipid nanoparticle (LNP) comprising a single stranded DNA (ssDNA), wherein the ssDNA:
- (a) encodes only one protein, which is a therapeutic protein,
- (b) is covalently closed,
- (c) does not form a double stranded structure longer than 40 base pairs,
- (d) is at least 200 nucleotides in length, and
- (e) does not comprise a protelomerase target sequence;
- wherein:
- (i) at least 95% by mass of total DNA in the composition is the ssDNA that is covalently closed;
- (ii) the pharmaceutical formulation is free of polypeptides;
- and
- (iii) the therapeutic protein is expressed when the ssDNA is introduced into a target cell.
A pharmaceutical formulation comprising:
- an LNP comprising a single stranded DNA (ssDNA), wherein the ssDNA:
- (a) encodes only one protein, which is a therapeutic protein,
- (b) is covalently closed,
- (c) does not form a double stranded structure longer than 100 base pairs,
- (d) is at least 200 nucleotides in length, and
- (e) does not comprise a protelomerase target sequence;
- wherein:
- (i) at least 95% by mass of total DNA in the composition is the ssDNA that is covalently closed;
- (ii) the pharmaceutical formulation is free of viral protein;
- and
- (iii) the therapeutic protein is expressed when the ssDNA is introduced into a target cell.
A pharmaceutical formulation comprising:
- an LNP comprising a single stranded DNA (ssDNA), wherein the ssDNA:
- (a) encodes a therapeutic protein,
- (b) is covalently closed,
- (c) does not form a double stranded structure longer than 100 base pairs,
- (d) is at least 200 nucleotides in length, and
- (e) does not comprise a protelomerase target sequence;
- wherein:
- (i) at least 95% by mass of total DNA in the composition is the ssDNA that is covalently closed;
- (ii) the pharmaceutical formulation is free of polypeptides;
- (iii) the therapeutic protein is expressed when the ssDNA is introduced into a target cell;
- and
- (iv) the ssDNA does not encode viral protein.
A pharmaceutical formulation comprising:
- an LNP comprising a single stranded DNA (ssDNA), wherein the ssDNA:
- (a) encodes a therapeutic protein,
- (b) is covalently closed,
- (c) does not comprise a region of intramolecular complementarity longer than 100 base pairs,
- (d) is at least 200 nucleotides in length, and
- (e) does not comprise a protelomerase target sequence;
- wherein:
- (i) at least 95% by mass of total DNA in the composition is the ssDNA that is covalently closed;
- (ii) the pharmaceutical formulation is free of polypeptides;
- (iii) the therapeutic protein is expressed when the ssDNA is introduced into a target cell;
- (iv) the therapeutic protein is selected from the group consisting of:
- a transcription factor, a chromatin remodeling factor, an antigen, a peptide, a hormone, an enzyme, an antibody, a receptor ligand, a receptor, a clotting factor, and a membrane protein;
- and
- (v) the ssDNA does not encode viral protein.
Dependencies are inferred from the claim text.