Granted Patent B2

US 12,209,250 · App. 16/741,827 · Granted Jan 28, 2025

Inventors: Ron Weiss (Newton, MA); Maria Hottelet Foley (Cambridge, MA); Jin Huh (Watertown, MA); Ross D. Jones (Cambridge, MA)

Assignee: Massachusetts Institute of Technology

C12N15/86A61K35/76A61K48/005C12N7/00C12N15/113C12N2710/16643C12N2800/10

Abstract

Claims (21)

1. An engineered Herpes Simplex Virus-1 (HSV-1) vector comprising a modified HSV-1 genome comprising non-functional deletions in both copies of the gene encoding Infected Cell Protein 4 (ICP4), non-functional deletions in both copies of the gene encoding Infected Cell Protein 0 (ICP0), a deletion in the gene encoding Virion Protein 16 (VP16) that results in a truncation of the VP16 protein at amino acid 422, and one or more deletions in one copy of the Latency Associated Transcript (LAT) region,

wherein a transcript of a LAT region is produced in an infected cell, and

wherein the HSV-1 genome comprises a nucleotide sequence having 95% sequence identity to the nucleotide sequence of SEQ ID NO: 1.

2. The engineered HSV-1 vector of claim 1 , wherein the modified HSV-1genome further comprises deletions in one or more genes encoding γ34.5, ICP6, ICP8, ICP27, ICP22, or ICP47.

3. The engineered HSV-1 vector of claim 2 , wherein one or more deletions render one or more of γ34.5, LAT, ICP27, ICP22, or ICP47 non-functional.

4. The engineered HSV-1 vector of claim 1 , wherein the modified HSV-1 genome is from HSV-1 strains F, 17, or KOS.

5. The engineered HSV-1 vector of claim 1 , further comprising one or more genetic circuits.

6. The engineered HSV-1 vector of claim 5 , wherein the one or more genetic circuits are up to 150 kb in length.

7. The engineered HSV-1 vector of claim 5 , wherein the one or more genetic circuits encodes an output molecule.

8. The engineered HSV-1 vector of claim 7 , wherein the output molecule is an HSV-1 protein, a therapeutic molecule, a diagnostic molecule, a functional molecule, or an inhibitor of innate immune response.

9. The engineered HSV-1 vector of claim 8 , wherein the inhibitor of innate immune response is an RNA interference (RNAi) molecule that targets an innate immune response component.

10. An isolated packaging cell comprising the engineered HSV-1 vector of claim 1 .

11. The isolated packaging cell of claim 10 , wherein the engineered HSV-1 vector is integrated into the genome of the isolated packaging cell.

12. The isolated packaging cell of claim 10 , wherein the isolated packaging cell produces at least 1 plaque forming units of HSV-1 viral particles.

13. The isolated packaging cell of claim 10 , wherein the isolated packaging cell is a U2OS cell.

14. An engineered HSV-1 viral particle comprising the engineered HSV-1 vector of claim 1 .

15. A method of treating a disease, the method comprising administering an effective amount of the engineered HSV-1 viral particle of claim 14 to a subject in need thereof, wherein the engineered HSV-1 vector of the engineered HSV-1 viral particle comprises a genetic circuit encoding a therapeutic molecule.

16. A method of diagnosing a disease, the method comprising administering an effective amount of the engineered HSV- 1 viral particle of claim 14 to a subject in need thereof, wherein the engineered HSV-1 vector of the engineered HSV-1 viral particle comprises a genetic circuit encoding a diagnostic molecule.

17. A method of delivering a genetic circuit into a cell, comprising contacting the cell with the engineered HSV-1 vector of claim 1 , wherein the engineered HSV-1 vector comprises a genetic circuit.

18. The engineered HSV-1 vector of claim 1 , wherein the modified HSV-1 genome comprises a nucleotide sequence having 99% sequence identity to the nucleotide sequence of SEQ ID NO: 1.

19. The engineered HSV-1 vector of claim 1 , wherein the modified HSV-1 genome comprises the nucleotide sequence of SEQ ID NO: 1.

Assignments (1)

Continuity (2)

References Cited (72)