Arrdc1-mediated microvesicles (armms) and uses thereof

Abstract

The invention provide isolated arrestin domain-containing protein 1 (ARRDC1)-mediated microvesicles (ARMMs). Methods for generating and for isolating ARMMs are also provided herein. ARMMs can be used to deliver agents, for example, nucleic acids (e.g., siRNAs, microRNAs, lincRNAs), proteins (e.g., transcription factors, chromatin modulators, kinases, phosphorylases, or recombinases), or small molecules to target cells in vitro and in vivo, and methods for such ARMM-mediated delivery are provided herein. Diagnostic and therapeutic methods using ARMMs are also described herein.

1 - 29 . (canceled) 
     
     
         30 . An ARRDC1 fusion protein, comprising
 an ARRDC1 protein or a fragment thereof, and   a polypeptide conjugated to the ARRDC1 protein or fragment thereof.   
     
     
         31 . The ARRDC1 fusion protein of  claim 30 , wherein the ARRDC1 fragment comprises a PSAP domain. 
     
     
         32 . A TSG101 fusion protein, comprising
 an TSG101 protein or a fragment thereof, and   a polypeptide conjugated to the TSG101 protein or fragment thereof.   
     
     
         33 . The TSG101 fusion protein of  claim 32 , wherein the TSG101 fragment comprises a UEV domain. 
     
     
         34 . The fusion protein of  claim 30 , wherein the polypeptide comprises a transcription factor, a transcriptional repressor, a fluorescent protein, a kinase, a phosphatase, a protease, a ligase, or a recombinase. 
     
     
         35 . The fusion protein of  claim 30 , wherein the polypeptide is conjugated to the ARRDC1 protein or the ARRDC1 protein fragment via a covalent bond. 
     
     
         36 . The fusion protein of  claim 30 , wherein the polypeptide is conjugated to the ARRDC1 protein or the ARRDC1 protein fragment via a linker. 
     
     
         37 . The fusion protein of  claim 36  wherein the linker is a cleavable linker. 
     
     
         38 . The fusion protein of  claim 37 , wherein the linker comprises a protease recognition site or a UV-cleavable moiety. 
     
     
         39 . The fusion protein of  claim 30 , wherein the polypeptide comprises a fluorescent protein. 
     
     
         40 - 86 . (canceled) 
     
     
         87 . The fusion protein of  claim 32 , wherein the polypeptide comprises a transcription factor, a transcriptional repressor, a fluorescent protein, a kinase, a phosphatase, a protease, a ligase, or a recombinase. 
     
     
         88 . The fusion protein of  claim 32 , wherein the polypeptide is conjugated to the TSG101 protein or the TSG101 protein fragment via a covalent bond. 
     
     
         89 . The fusion protein of  claim 32 , wherein the polypeptide is conjugated to the TSG101 protein or the TSG101 protein fragment via a linker. 
     
     
         90 . The fusion protein of  claim 89  wherein the linker is a cleavable linker. 
     
     
         91 . The fusion protein of  claim 90 , wherein the linker comprises a protease recognition site or a UV-cleavable moiety. 
     
     
         92 . The fusion protein of  claim 32 , wherein the polypeptide comprises a fluorescent protein. 
     
     
         93 . The fusion protein of  claim 92 , wherein the fluorescent protein is GFP, YFP, BFP, RFP, mCherry, or an enhanced or destabilized version thereof. 
     
     
         94 . A microvesicle-producing cell comprising a the fusion protein of  claim 30 . 
     
     
         95 . A microvesicle-producing cell comprising a recombinant expression construct encoding the fusion protein of  claim 30  under the control of a heterologous promoter. 
     
     
         96 . A method of delivering an agent to a target cell, the method comprising
 contacting the target cell with an isolated microvesicle comprising
 a lipid bilayer, 
 the fusion protein of  claim 30 , and 
 the agent.