Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

An enhanced packaging system for helper-dependent herpes simplex virus vectors

T A Stavropoulos1, C A Strathdee

  • 1Gene Therapy and Molecular Virology Group, The John P. Robarts Research Institute, London, Ontario, Canada N6A 5K8.

Journal of Virology
|August 8, 1998
PubMed
Summary

An enhanced packaging system using a modified helper virus and amplicon vector significantly increases herpes simplex virus (HSV) vector yields. This advancement improves the practical application of HSV amplicons for long-term gene expression.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

ASGT - Third Meeting. Stable gene delivery.

IDrugs : the investigational drugs journal·2005
Same author

Synergy between tetA and rpsL provides high-stringency positive and negative selection in bacterial artificial chromosome vectors.

Genomics·2001
Same author

Disease-associated mutations in SOD1 are impervious to dominant positive or negative effects.

Biochemical and biophysical research communications·2000
Same author

Expression of the tetA(C) tetracycline efflux pump in Escherichia coli confers osmotic sensitivity.

FEMS microbiology letters·2000
Same author

Biolistic-mediated interleukin 4 gene transfer prevents the onset of type 1 diabetes.

Human gene therapy·2000
Same author

Herpes simplex viral and amplicon vector-mediated gene transfer into glia and neurons in organotypic spinal cord and dorsal root ganglion cultures.

Molecular therapy : the journal of the American Society of Gene Therapy·2000

Area of Science:

  • Molecular Biology
  • Virology
  • Gene Therapy

Background:

  • Helper-dependent herpes simplex virus (HSV) vectors offer long-term gene expression potential.
  • Current packaging systems yield insufficient titers for practical applications.

Purpose of the Study:

  • To develop an enhanced second-generation packaging system for HSV amplicon vectors.
  • To increase the yield of packaged HSV amplicon vectors.

Main Methods:

  • Reverse engineered the helper virus into a single HSV-bacterial artificial chromosome (BAC) clone, deleting packaging elements.
  • Modified the HSV amplicon vector with a simian virus 40 origin of replication for independent replication.
  • Co-transfected 293T cells with the modified HSV-BAC helper virus and HSV amplicon.

Related Experiment Videos

Main Results:

  • Achieved a replication-proficient but packaging-defective HSV-1 genome.
  • Enabled HSV-independent vector replication for enhanced yield.
  • Obtained an eightfold increase in packaged-vector yield compared to the original system.
  • Ensured helper virus-free cell lysate.

Conclusions:

  • The enhanced packaging system significantly improves HSV amplicon vector production.
  • This system is suitable for gene therapy applications requiring high vector titers.
  • The modifications provide a more efficient and practical method for generating HSV vectors.