Transducible delivery of siRNA by dsRNA binding domain fusions to PTD/CPPS
AssigneeThe Regents of the University of California
InventorsSteven F. Dowdy, Jehangir S. Wadia, Bryan Meade, Akiko Eguchi
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Abstract
The disclosure provides fusion polypeptides and constructs useful in delivering anionically charged nucleic acid molecules including diagnostics and therapeutics to a cell or subject. The fusion constructs include a protein transduction domain and a nucleic acid binding domain, or a protein transduction domain and a nucleic acid that is coated with one or more nucleic acid binding domains sufficient to neutralize an anionic charge on the nucleic acid. Also provided are methods of treating disease and disorders such as cell proliferative disorders.
Claims30
A composition comprising:
- one or more double stranded RNA binding domains (DRBDs) in complex with an RNA interfering agent to form a DRBD-nucleic acid complex;
- and
- a plurality of protein transduction domains (PTD), wherein each of the plurality of PTDs is linked to another PTD, or one or more of the DRBDs of the DRBD-nucleic acid complex or both to render the DRBD-nucleic acid complex cationically charged at a physiological pH, and wherein each DRBD is linked to at least two PTDs.
A composition comprising:
- a) a first fusion polypeptide comprising:
- i) a plurality of protein transduction domains (PTDs), the PTDs comprising a membrane transport function;
- and
- ii) one or more double stranded RNA binding proteins (DRBDs);
- b) an RNA interfering agent, wherein the RNA interfering agent is anionically charged and interacts with the one or more DRBDs to form a protein-nucleic acid complex, and wherein the overall anionic charge of the fusion protein-nucleic acid complex is reduced relative to the nucleic acid alone;
- and
- c) a pharmaceutically acceptable carrier wherein the first fusion protein in complex with the RNA interfering agent has a net cationic charge at a physiological pH, and wherein each DRBD is linked to at least two PTDs.
A method of introducing an RNA interfering agent into a cell comprising:
- contacting the cell with a composition comprising one or more double stranded RNA binding domains (DRBDs) in complex with the RNA interfering agent to form a DRBD-nucleic acid complex and wherein the DRBD-nucleic acid complex is linked to a plurality of protein transduction domains (PTD), wherein each of the plurality of PTDs is linked to another PTD, or one or more of the DRBDs of the DRBD-nucleic acid complex or both to render the DRBD-nucleic acid complex cationically charged at a physiological pH, and wherein each DRBD is linked to at least two PTDs.
A kit comprising a vessel or vessels containing
- (a) one or more double stranded RNA binding domains (DRBDs) each linked to a plurality of protein transduction domains (PTDs), wherein each of the plurality of PTDs is linked to another PTD, or one or more of the DRBDs and wherein in each DRBD is linked to at least two PTDs;
- and
- (b) an RNA interfering agent.
A composition comprising:
- at least one fusion protein selected from the group consisting of:
- (i) a DRBD-single PTD fusion protein consisting of a double stranded RNA binding domain (DRBD) linked to one protein transduction domain;
- and
- (ii) a DRBD-multi-PTD fusion protein consisting of a DRBD and a plurality of PTDs linked to the DRBD;
- wherein the DRBD is in complex with an RNA interfering agent to form a DRBD-nucleic acid complex.
A composition consisting of:
- at least one fusion protein selected from the group consisting of:
- (i) a DRBD-single PTD fusion protein consisting of a double stranded RNA binding domain (DRBD) linked to one protein transduction domain;
- and
- (ii) a DRBD-multi-PTD fusion protein consisting of a DRBD and a plurality of PTDs linked to the DRBD;
- an RNA interfering agent in complex with the DRBD to form a DRBD-nucleic acid complex.
Dependencies are inferred from the claim text.