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Compositions and methods to treat latent viral infections

A virus infection and composition technology, which is applied in the field of compositions and methods for treating latent virus infection, and can solve problems such as not finding a treatment method.

Inactive Publication Date: 2017-08-01
THE BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIOR UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For these latent viruses, no treatment has been found to completely eradicate the infection

Method used

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  • Compositions and methods to treat latent viral infections
  • Compositions and methods to treat latent viral infections
  • Compositions and methods to treat latent viral infections

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0117] Example 1: Targeting EBV

[0118] Burkitt's lymphoma cell lines Raji, Namalwa and DG-75 were obtained from ATCC and cultured in RPMI 1640 supplemented with 10% FBS and PSA following ATCC recommendations. Human primary lung fibroblasts IMR-90 were obtained from Coriell and cultured in Advanced DMEM / F-12 supplemented with 10% FBS and PSA.

[0119] Chimeric guide RNAs (sgRNAs) driven by U6 promoters and ubiquitin promoters were obtained from Addgene as described by Cong L et al. (2013) Multiplex Genome Engineering Using CRISPR / CasSystems, Science 339:819-823. Plasmids composed of Cas9. The EGFP tag fused after the Cas9 protein allows selection of Cas9 positive cells ( Figure 1A ). We employed a modified chimeric guide RNA designed for more efficient Pol-III transcription and a more stable stem-loop structure (Chen B et al. (2013) Dynamic Imaging of Genomic Loci in Living Human Cells by an Optimized CRISPR / Cas System . Cell 155:1479-1491).

[0120] We obtained pX458 f...

Embodiment 2

[0141] Example 2: Targeting Hepatitis B Virus (HBV)

[0142] The methods and materials of the invention can be used to apply targeting endonucleases to specific genetic material, for example latent viral genomes such as hepatitis B virus (HBV). The invention also provides efficient and safe delivery of nucleic acids (eg, DNA plasmids) into target cells (eg, hepatocytes). In one embodiment, the methods of the invention use hydrodynamic gene delivery to target HBV.

[0143] Figure 10The HBV genome is shown. Annotation of the HBV genome (i.e., identification of important features of the genome) is preferably obtained and candidates for targeting by enzymatic degradation located in one of the features, e.g., viral origin of replication, terminal repeats, replication factors, are selected Binding sites, promoters, coding sequences and repeat regions.

[0144] HBV, the prototypical member of the hepadnavirus family, is a 42nm partially double-stranded DNA virus consisting of a ...

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Abstract

Viral infection is a persistent cause of human disease. Guided nuclease systems target the genomes of viral infections, rendering the viruses incapacitated.

Description

[0001] Cross References to Related Applications [0002] This application claims priority and benefit to U.S. Provisional Application Application No. 62 / 005,395, filed May 30, 2014, and U.S. Provisional Application Application No. 62 / 029,072, filed July 25, 2014, the entire contents of which are incorporated by reference This article. [0003] field of invention [0004] The present invention generally relates to compositions and methods for the selective treatment of viral infections using guided nuclease systems. Background technique [0005] Viral infection is a serious medical problem. There are various antiviral treatments, but they usually involve interrupting the replication cycle of the virus. Thus, a particularly difficult problem is latent viral infection, since there are no effective treatments capable of eradicating the virus from host cells. There is a continuing risk throughout life as latent infection can escape immune surveillance and reactivate the lytic ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/86C12N15/90C12N9/16C12N9/22C12N15/10C12N15/113
CPCC12N9/16C12N9/22C12N15/1133C12N15/86C12N15/907C12Y301/00C12N15/102A61K38/00C12N2330/51C12N2310/10C12N2810/60C12N2310/20A61P31/12A61P31/14A61P31/18A61P31/20A61P31/22A61P43/00Y02A50/30A61K47/6901A61K38/1761A61K48/005C12N2820/60
Inventor S·R·奎克王建斌
Owner THE BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIOR UNIV
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