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Generating Recombinant Avian Herpesvirus Vectors with CRISPR/Cas9 Gene Editing

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Herpes simplex virus type 1 (HSV-1)-derived amplicon vectors.

Matias E Melendez¹, Cornel Fraefel, Alberto L Epstein

¹Molecular Oncology Research Center, Barretos Cancer Hospital, 14784-400, Barretos, SP, Brazil.

Methods in Molecular Biology (Clifton, N.J.)

|March 28, 2014

Summary

Herpes simplex virus type 1 (HSV-1) amplicons offer safe gene transfer with large DNA capacity. New methods now provide helper-virus-free stocks for improved gene therapy applications.

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Area of Science:

Virology
Molecular Biology
Gene Therapy

Background:

Amplicons are defective, helper-dependent vectors derived from herpes simplex virus type 1 (HSV-1).
Their genomes lack protein-encoding viral sequences, ensuring host safety and cell non-toxicity.
They possess a large transgene capacity (up to 150-kb), suitable for delivering complete gene loci and regulatory elements.

Purpose of the Study:

To describe methodologies for generating helper-virus-free amplicon stocks.
To overcome limitations in amplicon vector development and application.

Main Methods:

Two distinct methodologies for producing amplicon stocks were developed.
These methods aim to eliminate or minimize contamination with toxic helper virus particles.

Main Results:

The developed methodologies allow for the generation of amplicon stocks free of helper particles or with only faint contamination.
This advancement significantly improves the utility of amplicon vectors for gene transfer.

Conclusions:

The described methods resolve a major hurdle in the development and application of HSV-1 amplicon vectors.
These improved amplicon stocks enhance their safety and efficacy for gene therapy and research.