Exosomes and methods of making and using the same
Provided herein are isolated exosomes. An exome can include a nonstructural (NSs) protein of a virus, a glycoprotein (Gn protein) of a virus, or a combination thereof. The virus is a member of the Order Bunyavirales, such as a member of the genus Phlebovirus . In one embodiment, the virus is Severe Fever with Thrombocytopenia Syndrome (SFTS) virus. Also provided are methods for making and using the exosomes described herein.
1. A method for making an exosome comprising
incubating an engineered cell under conditions suitable for production of an extracellular exosome, wherein the engineered cell comprises an exogenous polynucleotide encoding a nonstructural (NSs) protein of a virus that is a member of the Order Bunyavirales, or an active fragment thereof, wherein the extracellular exosome comprises marker LC3.
2. The method of claim 1 further comprising isolating the extracellular exosome.
3. The method of claim 1 wherein the virus is a member of the genus Phlebovirus.
4. The method of claim 3 wherein the member of the genus Phlebovirus is Severe fever with thrombocytopenia syndrome (SFTS) virus.
5. The method of claim 1 wherein the NSs protein has at least 80% similarity with SEQ ID NO:1.
6. The method of claim 1 wherein the number of extracellular exosomes produced by the cell is increased by at least 10-fold compared to the same cell that does not comprise the exogenous polynucleotide.
7. The method of claim 1 further comprising a glycoprotein (Gn protein) of a virus that is a member of the Order Bunyavirales, or an active fragment thereof.
8. The method of claim 1 wherein the extracellular exosome does not comprise a Severe fever with thrombocytopenia syndrome (SFTS) virus.
9. The method of claim 1 wherein the extracellular exosome further comprises an agent.
10. The method of claim 1 wherein the agent comprises a polynucleotide.
11. The method of claim 10 wherein the polynucleotide comprises mRNA, tRNA, rRNA, siRNA, microRNA, non-coding RNA, coding RNA, or DNA.
12. The method of claim 7 wherein the Gn protein has at least 80% similarity with SEQ ID NO:2.
13. The method of claim 7 wherein the Gn protein comprises SEQ ID NO:2.