IP Library › Granted Patent US 12,319,934
Granted Patent B2
US 12,319,934 · App. 18/129,292 · Granted Jun 3, 2025

Protein enriched microvesicles and methods of making and using the same

Inventors: Michael Haugwitz (Belmont, CA); Thomas Patrick Quinn (Sunnyvale, CA); Andrew Alan Farmer (Los Altos, CA); Montserrat Morell Fernández (Menlo Park, CA)
Assignee: Takara Bio USA, Inc.
C12N5/0602C07K14/005C07K14/43595C07K17/06C12N5/00C12N9/003C12N9/1241C12P21/02C12Y105/01003C07K2319/00C07K2319/70C12N2501/998C12N2760/00022C12N2760/20222
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Quick Facts
Patent No.
US 12,319,934
App. No.
18/129,292
Granted
Jun 3, 2025
Kind
B2
Abstract

Protein enriched micro-vesicles and methods of making and using the same are provided. Aspects of the methods include maintaining a cell having a membrane-associated protein comprising a first dimerization domain and a target protein having a second dimerization domain under conditions sufficient to produce a micro-vesicle from the cell, wherein the micro-vesicle includes the target protein. Also provided are cells, reagents and kits that find use in making the micro-vesicles, as well as methods of using the micro-vesicles, e.g., in research and therapeutic applications.

Claims (18)

1. A method of introducing a target protein into a cell, the method comprising:

contacting the cell with a micro-vesicle comprising:

(a) a chimeric membrane-associated protein comprising a membrane-associated domain and a first dimerization domain, wherein the membrane-associated domain and the first dimerization domain are heterologous; and

(b) a chimeric target protein comprising a second dimerization domain, wherein the target protein is not membrane-associated in the absence of the chimeric membrane-associated protein;

wherein the first and second dimerization domains are specifically bound to each other in a dimerized complex or the first and second dimerization domains are bound to each other by a single dimerization mediator;

under conditions sufficient for the micro-vesicle to fuse with the cell and introduce the target protein into the cell.

2. The method according to claim 1 , wherein the chimeric membrane-associated protein and chimeric the target protein are present in a dimerized complex.

3. The method according to claim 2 , wherein the first and second dimerization domains are specifically bound to each other in the dimerized complex.

4. The method according to claim 2 , wherein first and second dimerization domains are bound to each other by a dimerization mediator.

5. The method according to claim 4 , wherein the dimerization mediator is a modifiable dimerization mediator.

6. The method according to claim 5 , wherein the method further comprises applying a stimulus to the micro-vesicle to modify the dimerization mediator to dissociate the complex of the chimeric target protein and the chimeric membrane-associated protein.

7. The method according to claim 2 , further comprising contacting the micro-vesicle with a dimerization disruptor to dissociate the chimeric target protein from the chimeric membrane-associated protein.

8. The method according to claim 1 , wherein the micro-vesicle further comprises a second chimeric target protein comprising a third dimerization domain.

9. The method according to claim 1 , wherein the chimeric target protein comprises a therapeutic protein.

10. The method according to claim 1 , wherein the micro-vesicle exhibits tropism for the cell.

11. The method according to claim 1 , wherein the cell is a human cell.

12. The method according to claim 1 , wherein the cell is in vitro.

13. The method according to claim 1 , wherein the cell is in vivo.

Assignments (2)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Mar 31, 2023
From: HAUGWITZ, MICHAEL; QUINN, THOMAS PATRICK; FARMER, ANDREW ALAN; FERNÁNDEZ, MONTSERRAT MORELL
To: CLONTECH LABORATORIES, INC.
Reel/Frame 063189/0155 →
CHANGE OF NAME Recorded Mar 31, 2023
From: CLONTECH LABORATORIES, INC.
To: TAKARA BIO USA, INC.
Reel/Frame 063227/0397 →
Continuity (6)
Division 16999399 · Aug 21, 2020
Continuation 15415754 · Jan 25, 2017
Division 14278714 · May 15, 2014
Provisional Application 61872115 · Aug 30, 2013
Provisional Application 61833880 · Jun 11, 2013
Related Publication 20230235278A1 · Jul 27, 2023
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