Abstract: 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.

Abstract: Compositions and methods for delivering non-anionic polynucleotide analog cargoes to the cytosolic/nuclear compartment of eukaryotic cells via a synthetic peptide shuttle agent are described herein. The non-anionic polynucleotide analog cargoes may be charge-neutral or cationic, and the synthetic peptide shuttle agent is 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 is not covalently linked, or linked in a cleavable manner under physiological conditions, to the non-anionic polynucleotide analog cargoes. The non-anionic polynucleotide analog cargo may be a charge-neutral or cationic antisense synthetic oligonucleotide (ASO) that hybridizes to an intracellular target RNA for gene expression modification.

Abstract: The present description relates to methods for delivering polypeptide cargos from an extracellular space to the cytosol and/or nucleus of a target eukaryotic cell. The methods involve contacting the cell with the polypeptide cargo in the presence of a peptide shuttle agent at a concentration sufficient to increase the polypeptide cargo's transduction efficiency. Also described here are parameters that may be used in the rational design of such synthetic peptide shuttle agents, peptide shuttle agents that satisfy one or more of these design parameters, as well as methods and compositions relating to the use of the synthetic peptide shuttle agents for delivery of a variety of polypeptide cargos (such as transcription factors, antibodies, CRISPR-associated nucleases and functional genome editing complexes) from an extracellular space to the cytosol and/or nucleus of target eukaryotic cells. Applications and targets for genome-editing NK cells for improved immunotherapy are also described.

Abstract: The present description relates to methods for delivering polypeptide cargos from an extracellular space to the cytosol and/or nucleus of a target eukaryotic cell. The methods involve contacting the cell with the polypeptide cargo in the presence of a peptide shuttle agent at a concentration sufficient to increase the polypeptide cargo's transduction efficiency. Also described here are parameters that may be used in the rational design of such synthetic peptide shuttle agents, peptide shuttle agents that satisfy one or more of these design parameters, as well as methods and compositions relating to the use of the synthetic peptide shuttle agents for delivery of a variety of polypeptide cargos (such as transcription factors, antibodies, CRISPR-associated nucleases and functional genome editing complexes) from an extracellular space to the cytosol and/or nucleus of target eukaryotic cells. Applications and targets for genome-editing NK cells for improved immunotherapy are also described.

Abstract: Described herein are methods, compositions, kits and synthetic peptide shuttle agents relating to the transduction of proteinaceous and/or non-proteinaceous cargoes. The method generally comprises contacting target eukaryotic cells with a non-proteinaceous cargo and a concentration of a synthetic peptide shuttle agent sufficient to increase the transduction efficiency of the non-proteinaceous cargo, as compared to in the absence of said synthetic peptide shuttle agent. In embodiments, the non-proteinaceous cargo may be a drug, such as a small molecule drug, for treating a disease. In other embodiments, novel synthetic peptide shuttle agents having transduction activity for proteinaceous and/or non-proteinaceous cargoes are described, as well as the use of propidium iodide or other membrane-impermeable fluorescent DNA intercalating agent as a surrogate cargo for selecting versatile synthetic peptide shuttle agents having transduction activity for both proteinaceous and non-proteinaceous cargoes.

Abstract: The present description relates to methods for delivering polypeptide cargos from an extracellular space to the cytosol and/or nucleus of a target eukaryotic cell. The methods involve contacting the cell with the polypeptide cargo in the presence of a peptide shuttle agent at a concentration sufficient to increase the polypeptide cargo's transduction efficiency. Also described here are parameters that may be used in the rational design of such synthetic peptide shuttle agents, peptide shuttle agents that satisfy one or more of these design parameters, as well as methods and compositions relating to the use of the synthetic peptide shuttle agents for delivery of a variety of polypeptide cargos (such as transcription factors, antibodies, CRISPR-associated nucleases and functional genome editing complexes) from an extracellular space to the cytosol and/or nucleus of target eukaryotic cells. Applications and targets for genome-editing NK cells for improved immunotherapy are also described.

Abstract: The present description relates to synthetic peptides useful for increasing the transduction efficiency of polypeptide cargos to the cytosol of target eukaryotic cells. More specifically, the present description relates to synthetic peptides and polypeptide-based shuttle agents comprising an endosome leakage domain (ELD) operably linked to a cell penetrating domain (CPD), or an ELD operably linked to a histidine-rich domain and a CPD. Compositions, kits, methods and uses relating to same are also described.

Abstract: The present description relates to synthetic peptides useful for increasing the transduction efficiency of polypeptide cargos to the cytosol of target eukaryotic cells. More specifically, the present description relates to synthetic peptides and polypeptide-based shuttle agents comprising an endosome leakage domain (ELD) operably linked to a cell penetrating domain (CPD), or an ELD operably linked to a histidine-rich domain and a CPD. Compositions, kits, methods and uses relating to same are also described.

Abstract: The present description relates to methods for delivering polypeptide cargos from an extracellular space to the cytosol and/or nucleus of a target eukaryotic cell. The methods involve contacting the cell with the polypeptide cargo in the presence of a peptide shuttle agent at a concentration sufficient to increase the polypeptide cargo's transduction efficiency. Also described here are parameters that may be used in the rational design of such synthetic peptide shuttle agents, peptide shuttle agents that satisfy one or more of these design parameters, as well as methods and compositions relating to the use of the synthetic peptide shuttle agents for delivery of a variety of polypeptide cargos (such as transcription factors, antibodies, CRISPR-associated nucleases and functional genome editing complexes) from an extracellular space to the cytosol and/or nucleus of target eukaryotic cells. Applications and targets for genome-editing NK cells for improved immunotherapy are also described.

Abstract: The present description relates to methods for delivering polypeptide cargos from an extracellular space to the cytosol and/or nucleus of a target eukaryotic cell. The methods involve contacting the cell with the polypeptide cargo in the presence of a peptide shuttle agent at a concentration sufficient to increase the polypeptide cargo's transduction efficiency. Also described here are parameters that may be used in the rational design of such synthetic peptide shuttle agents, peptide shuttle agents that satisfy one or more of these design parameters, as well as methods and compositions relating to the use of the synthetic peptide shuttle agents for delivery of a variety of polypeptide cargos (such as transcription factors, antibodies, CRISPR-associated nucleases and functional genome editing complexes) from an extracellular space to the cytosol and/or nucleus of target eukaryotic cells. Applications and targets for genome-editing NK cells for improved immunotherapy are also described.

Abstract: The present description relates to synthetic peptides useful for increasing the transduction efficiency of polypeptide cargos to the cytosol of target eukaryotic cells. More specifically, the present description relates to synthetic peptides and polypeptide-based shuttle agents comprising an endosome leakage domain (ELD) operably linked to a cell penetrating domain (CPD), or an ELD operably linked to a histidine-rich domain and a CPD. Compositions, kits, methods and uses relating to same are also described.

Abstract: The present description relates to synthetic peptides useful for increasing the transduction efficiency of polypeptide cargos to the cytosol of target eukaryotic cells. More specifically, the present description relates to synthetic peptides and polypeptide-based shuttle agents comprising an endosome leakage domain (ELD) operably linked to a cell penetrating domain (CPD), or an ELD operably linked to a histidine-rich domain and a CPD. Compositions, kits, methods and uses relating to same are also described.

Abstract: The present description relates to synthetic peptides useful for increasing the transduction efficiency of polypeptide cargos to the cytosol of target eukaryotic cells. More specifically, the present description relates to synthetic peptides and polypeptide-based shuttle agents comprising an endosome leakage domain (ELD) operably linked to a cell penetrating domain (CPD), or an ELD operably linked to a histidine-rich domain and a CPD. Compositions, kits, methods and uses relating to same are also described.

Abstract: A nectar feeder for birds. The nectar feeder has a container for storing nectar feed. A base tray is attached to the container and connected in fluid communication with the container. A float is positioned between the container and the base tray. A ball is positioned on the float. The ball regulates the fluid communication between the container and the base tray. In operation, the nectar feed flows through the container and into the base tray until the float rises to a level where the ball prevents fluid from flowing into the base tray.

Abstract: The present invention is a nectar feeder for birds. The nectar feeder has a container for storing nectar feed. A base tray is connected in fluid communication with the container. A float is positioned between the container and the base tray. In operation, the nectar feed flows through the container and into the base tray until the float rises to a level to prevent fluid from flowing into the base tray.

Abstract: The present invention is a nectar feeder for birds. The nectar feeder has a container for storing nectar feed. A base tray is attached to the container and connected in fluid communication with the container. A float is positioned between the container and the base tray. A ball is positioned on the float. The ball regulates the fluid communication between the container and the base tray. In operation, the nectar feed flows through the container and into the base tray until the float rises to a level where the ball prevents fluid from flowing into the base tray.

Abstract: The present invention is a nectar feeder for birds. The nectar feeder has a container for storing nectar feed. A base tray is attached to the container and connected in fluid communication with the container. A float is positioned between the container and the base tray. A ball is positioned on the float. The ball regulates the fluid communication between the container and the base tray. In operation, the nectar feed flows through the container and into the base tray until the float rises to a level where the ball prevents fluid from flowing into the base tray.

Abstract: The present invention is a nectar feeder for birds. The nectar feeder has a container for storing nectar feed. A base tray is attached to the container and connected in fluid communication with the container. A float is positioned between the container and the base tray. A ball is positioned on the float. The ball regulates the fluid communication between the container and the base tray. In operation, the nectar feed flows through the container and into the base tray until the float rises to a level where the ball prevents fluid from flowing into the base tray.