Non-integrating viral delivery system and methods related thereto

Inventors

Pauza, Charles David • Lahusen, Tyler

Assignees

American Gene Technologies International Inc

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.

Core Innovation

The invention relates to a non-integrating viral delivery system comprising a viral carrier containing a defective integrase gene and a heterologous viral episomal origin of DNA replication. The heterologous viral episomal origin of DNA replication comprises a sequence having at least 95% sequence identity with SEQ ID NO: 1, and the system includes a first sequence encoding at least one initiator protein specific for the heterologous viral episomal origin of DNA replication, where expression of the first sequence is inducible.

The inducible initiator protein regulates expression of a second sequence encoding at least one gene, gene product, shRNA, siRNA, miRNA, or other RNA of interest. In described embodiments, the system uses papillomavirus episomal elements, including HPV16 long control region (LCR) and LCR fragments, together with initiator proteins E1 and/or E2 and/or EBNA-1.

The control concept includes creating an “off” switch by withdrawing inducer so that initiator expression is controlled. The described payload capabilities include therapeutic or diagnostic targets such as VEGF, anti-HER2/anti-EGFR antibodies, and RNA molecules such as CCR5 miRNA (miR-CCR5), including shRNA/siRNA/other RNA of interest.

The document further describes testing of episome replication and payload expression with inducible E1/E2/EBNA-1 systems and different HPV16 LCR fragment sizes, including readouts such as GFP/mCherry, VEGF ELISA, antibody detection, and CCR5 knockdown measurements.

Claims Coverage

The independent claim requires a non-integrating viral delivery system with four inventive features: a viral carrier containing a defective integrase gene, a heterologous viral episomal origin of DNA replication with at least 95% identity to SEQ ID NO: 1, an inducible initiator protein sequence specific for that origin, and a second sequence encoding a gene or RNA of interest whose expression is regulated by the initiator. No other independent claims are provided in the supplied claim list.

Overall, the claim coverage centers on a non-integrating viral delivery system that combines a defective integrase gene carrier, a heterologous viral episomal replication origin of at least 95% identity to SEQ ID NO: 1, and an inducible, origin-specific initiator protein that regulates a payload sequence encoding a gene, gene product, or RNA of interest.

Stated Advantages

Documented Applications

No documented applications found