IP Library › Granted Patent US 10,907,130
Granted Patent B2
US 10,907,130 · App. 15/524,963 · Granted Feb 2, 2021

Methods and materials for producing recombinant viruses in eukaryotic microalgae

Inventor: Brian K. Kaspar (Westerville, OH)
Assignee: RESEARCH INSTITUTE AT NATIONWIDE CHILDREN'S HOSPITAL
C12N1/12C12N7/00C12N15/86C12N2750/14122C12N2750/14143C12N2750/14151C12N2800/22
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 10,907,130
App. No.
15/524,963
Granted
Feb 2, 2021
Kind
B2
Abstract

The present invention is directed to methods and materials for producing recombinant viruses. In particular, methods and materials are provided for producing recombinant viruses in eukaryotic microalgae such as Chlamydomonas reinhardtii . Recombinant adeno-associated viruses are examples of recombinant viruses produced according to the invention.

Claims (24)

1. A eukaryotic microalgae that produces a recombinant adeno-associated virus (rAAV).

2. The eukaryotic microalgae of claim 1 wherein the eukaryotic microalgae is Chlamydomonas reinhardtii, Chlorella vulgaris, Chlorella ellipsoidea, Chlorella sorokiniana, Chlorella kessleri, Volvox carteri, Dunaliella salina, Ostreococcus tauri, Phaeodactylum tico, Gonium pectoral , and Cyanidiosschyzon merolae.

3. The eukaryotic microalgae of claim 1 or 2 wherein the eukaryotic microalgae is Chlamydomonas reinhardtii.

4. The eukaryotic microalgae of claim 1 , wherein the microalgae is transformed with a polynucleotide construct comprising (i) a polynucleotide sequence encoding Rep 78 protein, Rep 52 protein, VP1 protein or VP2/3 protein or (ii) one or more polynucleotide sequences encoding Rep78 protein, Rep52 protein, VP1 protein and VP2/VP3 protein or (iii) one or more polynucleotide sequences encoding Rep78 protein, Rep52 protein, VP1 protein, VP2 protein and VP3 protein.

5. The eukaryotic microalgae of claim 4 wherein the construct further comprises an AAV inverted terminal repeat (ITR) and 3′ AAV ITR, and helper functions for generating a productive AAV infection.

6. The eukaryotic microalgae of claim 1 , wherein the microalgae is transformed with one or more polynucleotide constructs wherein each polynucleotide construct comprises at least one polynucleotide sequence encoding Rep 78 protein, Rep 52 protein, VP1 protein, VP2/3 protein, VP2 protein or VP3 protein.

7. The eukaryotic microalgae of claim 6 wherein the construct further comprises an AAV inverted terminal repeat (ITR) and 3′ AAV ITR, and helper functions for generating a productive AAV infection.

8. The eukaryotic microalgae of claim 6 , wherein the recombinant AAV is AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12 or AAV13.

9. The eukaryotic microalgae of claim 6 , wherein the recombinant AAV is AAV9.

10. The eukaryotic microalgae of claim 1 , wherein the recombinant AAV is AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12 or AAV13.

11. The eukaryotic microalgae of claim 1 , wherein the recombinant AAV is AAV9.

12. A method of producing a recombinant virus, wherein the recombinant virus is a recombinant adeno-associated virus (rAAV), comprising the steps of growing an eukaryotic microalgae producing the recombinant virus.

13. The method of claim 12 wherein the eukaryotic microalgae is transformed with a polynucleotide sequence expressing the recombinant virus.

14. The method of claim 12 further comprising the step of purifying the recombinant virus.

15. The method of claim 12 , wherein the eukaryotic microalgae is Chlamydomonas reinhardtii, Chlorella vulgaris, Chlorella ellipsoidea, Chlorella sorokiniana, Chlorella kessleri, Volvox carteri, Dunaliella saline, Ostreococcus tauri, Phaeodactylum tico, Gonium pectoral , and Cyanidiosschyzon merolae.

16. The method of claim 12 , wherein the eukaryotic microalgae is Chlamydomonas reinhardtii.

17. The method of claim 12 , wherein the recombinant AAV is AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12 or AAV13.

18. The method of claim 12 , wherein the recombinant AAV is AAV9.

19. The method of claim 12 , wherein the eukaryotic microalgae is transformed with a polynucleotide construct comprising (i) a polynucleotide nucleotide sequence encoding Rep 78 protein, Rep 52 protein, VP1 protein or VP2/3 protein or (ii) one or more polynucleotide sequences encoding Rep78 protein, Rep52 protein, VP1 protein and VP2/VP3 protein or (iii) one or more polynucleotide sequences encoding Rep78 protein, Rep52 protein, VP1 protein, VP2 protein and VP3 protein.

20. The method of claim 19 wherein the construct further comprises an AAV inverted terminal repeat (ITR) and 3′ AAV ITR, and helper functions for generating a productive AAV infection.

21. The method of claim 12 , wherein the eukaryotic microalgae is transformed with one or more polynucleotide constructs wherein each construct comprises at least one polynucleotide sequence encoding Rep 78 protein, Rep 52 protein, VP1 protein, VP2/3 protein, VP2 protein or VP3 protein.

22. The method of claim 21 wherein the construct further comprises an AAV inverted terminal repeat (ITR) and 3′ AAV ITR, and helper functions for generating a productive AAV infection.

23. The method of claim 21 , wherein the recombinant AAV is AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12 or AAV13.

24. The method of claim 21 , wherein the recombinant AAV is AAV9.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jul 28, 2017
From: KASPAR, BRIAN K.
To: RESEARCH INSTITUTE AT NATIONWIDE CHILDREN'S HOSPITAL
Reel/Frame 043133/0769 →
Continuity (2)
Provisional Application 62075408 · Nov 5, 2014
Related Publication 20180282684A1 · Oct 4, 2018