Phase changing formulations of nucleic acid payloads
The present invention is based, at least in part, upon discovery of a process for identifying phase changing peptides. Such phase changing peptides are capable of enhancing in vitro and in vivo delivery of oligonucleotides (e.g., dsRNAs) in lipidic, vesicular, micellar and/or naked oligonucleotide formulations.
1. A method for reducing expression of a target gene in a cell of a subject, comprising:
contacting said cell with a formulation comprising a DsiRNA and a phase changing peptide, wherein said peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs.: 1-50, in an amount effective to reduce expression of said target gene in said cell in comparison to a reference dsRNA.
2. The method of claim 1 , wherein said DsiRNA is a DsiRNA of 25-30 nucleotides.
3. The method of claim 1 , wherein said DsiRNA is a DsiRNA of 30-70 nucleotides.
4. The method of claim 1 , wherein said formulation is selected from the group consisting of a lipid formulation and a polymer formulation.
5. The method of claim 1 , wherein said formulation is selected from the group consisting of a vesicle based formulation and a micelle based formulation.
6. The method of claim 1 , wherein said formulation comprises free DsiRNA in solution.
7. The method of claim 1 , wherein said peptide further comprises a linker comprising one or more amino acids.
8. The method of claim 7 , wherein said linker comprises a cysteine residue.
9. The method of claim 7 , wherein said linker comprises a structure selected from the group consisting of a Citruline-Valine (Cit-Val) structure, a Valine-Citruline (Val-Cit) structure, and a repeat thereof.
10. The method of claim 7 , wherein said linker is attached to said DsiRNA.
11. The method of claim 10 , wherein said peptide further comprises spacer amino acids.
12. The method of claim 11 , wherein said spacer amino acids comprise Glycine and/or Serine.
13. The method of claim 1 , wherein said formulation further comprises PEG.
14. The method of claim 1 , wherein said peptide is attached to PEG.
15. The method of claim 1 , wherein said peptide is conjugated to said DsiRNA.
16. The method of claim 15 , wherein said conjugated peptide is in the aqueous phase of the formulation.
17. The method of claim 15 , wherein said conjugated peptide is in the lipid or polymer phase of the formulation.
18. The method of claim 1 , wherein said formulation is a lipid nanoparticle.