Exosome transfer of nucleic acids to cells

Abstract

Methods for introducing nucleic acids to cells via exosomes for use in gene modulation and therapy, such as for gene silencing and to introduce genetic material into cells to compensate for abnormal genes or to induce or repress a process in the recipient cell.

1 . A method of transferring genetic material, comprising:
 a) providing exosomes containing selected genetic material; and   b) transferring the selected genetic material from the exosomes to recipient cells.   
     
     
         2 . The method of  claim 1 , wherein the selected genetic material is selected from the group consisting of mRNA, tRNA, rRNA, siRNA, regulating RNA, non-coding and coding RNA, DNA fragments, and DNA plasmids. 
     
     
         3 . The method of  claim 1 , wherein use of recipient cells are selected from the group consisting of cells from recipients having inherited diseases in hematopoietic, non-hematopoietic, stem cells, and organs. 
     
     
         4 . The method of  claim 1 , wherein the selected material comprises nucleic acids and the recipient cells comprise CD4 T-cells for treatment of HIV infected T-cells. 
     
     
         5 . The method of  claim 1 , wherein the selected genetic material is transferred to treat diseases, induce or repress cell death, change cellular ageing, induce tolerance, re-direct existing immune responses, or change intracellular activity or cellular behaviour. 
     
     
         6 . The method of  claim 1 , wherein providing the exosomes further comprises a) producing exosomes lacking genetic material; and b) adding selected genetic material to the exosomes lacking genetic material. 
     
     
         7 . The method of  claim 6 , wherein the selected genetic material is selected from the group consisting of mRNA, tRNA, rRNA, siRNA, regulating RNA, non-coding and coding RNA, DNA fragments, and DNA plasmids. 
     
     
         8 . The method of  claim 7 , wherein the selected genetic material is added to the exosomes lacking genetic material using a method selected from the group consisting of transformation, transfection and microinjection of the selected genetic material into exosomes. 
     
     
         9 . The method of  claim 1 , wherein the exosomes are produced by a method selected from the group consisting of isolation from a selected type of donor cell; isolation from a person with a particular disease or condition; and isolation from a genetically modified donor cell. 
     
     
         10 . The method of  claim 9 , further comprising genetically modifying the donor cell to erase production of a nucleic acid, or to up-regulate or down-regulate production of a nucleic acid. 
     
     
         11 . The method of  claim 1 , further comprising introducing the selected genetic material into the exosomes by introducing the selected genetic material into donor cells followed by intracellular translocation of the genetic material into the exosomes. 
     
     
         12 . The method of  claim 1 , further comprising introducing the selected genetic material into the exosomes using a method selected from the group consisting of transformation, transfection and microinjection of the selected genetic material into the exosomes. 
     
     
         13 . The method of  claim 1 , wherein the genetic material is transferred to the recipient cells by a method selected from the group consisting of addition of the exosomes to cell cultures in vitro, intravenous injection of the exosomes, in vivo administration, and administration targeted to particular cells in the body. 
     
     
         14 . The method of  claim 1 , wherein the exosomes containing selected genetic material are taken from a patient with a malignant disease; and are further processed to contain particular genetic constructs; and then are reintroduced to the patient. 
     
     
         15 . A composition of isolated exosomes, wherein the isolated exosomes contain one or more selected nucleic acid constructs, and wherein the isolated exosomes do not contain any substantial amount of nucleic acids other than the selected nucleic acid constructs. 
     
     
         16 . The composition of  claim 15 , wherein the selected nucleic acid constructs are selected from the group consisting of mRNA, tRNA, rRNA, siRNA, microRNA, regulating RNA, non-coding and coding RNA, DNA fragments, and DNA plasmids. 
     
     
         17 . The composition of  claim 1 , wherein the isolated exosomes containing one or more selected nucleic acid constructs are isolated from donor cells, wherein the donor cells express the nucleic acid constructs or have been transfected with the nucleic acid constructs. 
     
     
         18 . The composition of  claim 1 , wherein the exosomes were isolated from a specific donor cell, or from a person with a particular disease or condition, or from a genetically modified donor cell. 
     
     
         19 . The composition of  claim 1 , wherein the exosomes were obtained from (i) tumor tissue, (ii) intestinal epithelial cells, lymphocytes, mast cells or dendritic cells, (iii) in vitro growing cells, (iv) cell culture supernatant, (v) body fluid, or (vi) any combination of (i) to (v). 
     
     
         20 . A recipient cell, wherein the recipient cell comprises a selected nucleic acid construct introduced to the recipient cell via an isolated exosome of  claim 1 .