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AssigneeFeldan Bio Inc.
InventorsDavid GUAY, Stéphanie HALLÉE, Xavier BARBEAU, Thomas DEL'GUIDICE, Jean-Pascal LEPETIT-STOFFAES
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Compositions and methods for delivering nucleoprotein cargos such as Cas9-RNP genome editing and ABE-Cas9-RNP base editing complexes to the cytosolic/nuclear compartment of eukaryotic cells via synthetic peptide shuttle agents are described herein. Also described herein are shortened synthetic peptide shuttle agents having a length of less than 20 amino acids having defined geometries associated with cargo transduction activity. The synthetic peptide shuttle agents are peptides comprising an amphipathic alpha-helical motif having both a positively-charged hydrophilic outer face and a hydrophobic outer face, wherein the synthetic peptide shuttle agent is independent from or is not covalently linked to the cargoes. Shuttle agents engineered for increased resistance to inhibition by nucleoproteins and/or extracellular DNA/RNA are also described herein.
A composition comprising a nucleoprotein cargo for intracellular delivery and a synthetic peptide shuttle agent that is independent from, or is not covalently linked to, said nucleoprotein cargo, the synthetic peptide shuttle agent being a peptide comprising an amphipathic alpha-helical motif having both a positively-charged hydrophilic outer face and a hydrophobic outer face, wherein synthetic peptide shuttle agent increases cytosolic/nuclear delivery of said nucleoprotein cargo in eukaryotic cells as compared to in the absence of the synthetic peptide shuttle agent.
The composition of any one of 18 to 29, wherein the synthetic peptide shuttle agent is at least 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 amino acids in length.
A method for cargo transduction, the method comprising contacting target eukaryotic cells with said cargo and a concentration of a synthetic peptide shuttle agent sufficient to increase the transduction efficiency of said cargo, as compared to in the absence of said synthetic peptide shuttle agent, wherein said synthetic peptide shuttle agent is a peptide less than 20 amino acids in length, and wherein the shuttle agent and cargo are not covalently bound at the time of transduction across the plasma membrane.
A synthetic peptide shuttle agent for use in, or suitable for use in, the delivery of non-anionic cargoes across mucus-producing membranes (e.g., airway epithelium), the synthetic peptide shuttle agent comprising or consisting essentially of a central core amphipathic alpha helical region having shuttle agent activity, flanked N- and C-terminally by flexible linker domains, wherein one or both of the flexible linker domains comprises or consists essentially of a sufficient number of non-cationic hydrophilic residues such that cargo transduction activity across mucus-producing membranes of the synthetic peptide shuttle agent is increased relative to that of the central core amphipathic alpha helical region lacking the flexible linker domains.
Dependencies are inferred from the claim text.