Non-integrating viral delivery system and methods related thereto
AssigneeAmerican Gene Technologies International Inc.
InventorsCharles David Pauza, Tyler Lahusen
Life of this patent
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Abstract
A non-integrating viral delivery system is disclosed. The system includes a viral carrier, wherein the viral carrier contains a defective integrase gene; a heterologous viral episomal origin of replication; a sequence encoding at least one initiator protein specific for the heterologous viral episomal origin of replication, wherein expression of the sequence encoding the at least one initiator protein specific for the heterologous viral episomal origin of DNA replication is inducible; and at least one gene, gene product, shRNA, siRNA, miRNA, or other RNA of interest.
Claims36
A non-integrating viral delivery system, the system comprising:
- (a) a viral carrier, wherein the viral carrier contains a defective integrase gene;
- (b) a heterologous viral episomal origin of DNA replication;
- (c) a sequence encoding at least one initiator protein specific for the heterologous viral episomal origin of DNA replication, wherein expression of the sequence encoding the at least one initiator protein specific for the heterologous viral episomal origin of DNA replication is inducible;
- and
- (d) at least one gene, gene product, shRNA, siRNA, miRNA, or other RNA of interest.
A method of expressing at least one gene, gene product, shRNA, siRNA, miRNA, or other RNA of interest in a cell, the method comprising:
- contacting the cell with an effective amount of a non-integrating viral delivery system, wherein the system comprises:
- i. a viral carrier, wherein the viral carrier contains a defective integrase gene;
- ii. a heterologous viral episomal origin of DNA replication;
- iii. a sequence encoding at least one initiator protein specific for the heterologous viral episomal origin of DNA replication, wherein expression of the sequence encoding the at least one initiator protein specific for the heterologous viral episomal origin of DNA replication is inducible;
- and
- iv. at least one gene, gene product, shRNA, siRNA, miRNA, or other RNA of interest.
A method of expressing at least one gene, gene product, shRNA, siRNA, miRNA, or other RNA of interest in a subject in need thereof, the method comprising:
- administering to the subject in need thereof an effective amount of a non-integrating viral delivery system, wherein the system comprises:
- i. a viral carrier, wherein the viral carrier contains a defective integrase gene;
- ii. a heterologous viral episomal origin of replication;
- iii. a sequence encoding at least one initiator protein specific for the heterologous viral episomal origin of DNA replication, wherein expression of the sequence encoding the at least one initiator protein specific for the heterologous viral episomal origin of DNA replication is inducible;
- and
- iv. at least one gene, gene product, shRNA, siRNA, miRNA, or other RNA of interest.
A method of selecting an optimized non-integrating viral delivery system, the method comprising:
- selecting a level of basal expression, wherein when level X is selected, a corresponding Y is selected, wherein Y corresponds to a heterologous viral episomal origin of DNA replication selected to be incorporated into the non-integrating viral delivery system, whereby:
- when X=low;
- Y comprises SEQ ID NO:
- 1 or Frag1 (SEQ ID NO:
- 2);
- and
- when X=moderate;
- Y comprises Frag2 (SEQ ID NO:
- 3), Frag3 (SEQ ID NO:
- 4), or Frag4 (SEQ ID NO:
- 5) of the LCR of HPV16.
Dependencies are inferred from the claim text.