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

Replication-competent retrovirus produced by a 'split-function' third generation amphotropic packaging cell line

H Chong1, R G Vile

  • 1Imperial Cancer Research Fund Laboratory of Cancer Gene Therapy, Rayne Institute, St Thomas' Hospital, London, UK.

Gene Therapy
|July 1, 1996
PubMed
Summary
This summary is machine-generated.

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

The blister that got out of hand.

Clinical and experimental dermatology·2021
Same author

Effect of isoflurane on myocardial ischemia-reperfusion injury through the p38 MAPK signaling pathway.

European review for medical and pharmacological sciences·2019
Same author

Multiple perioral homogenous blue macules.

Clinical and experimental dermatology·2018
Same author

Acrodermatitis acidaemica.

Clinical and experimental dermatology·2018
Same author

Lichen planus in a patient treated with pembrolizumab for metastatic malignant melanoma.

Clinical and experimental dermatology·2017
Same author

Necrobiotic xanthogranuloma treated with lenalidomide.

Clinical and experimental dermatology·2017
Same journal

Improving the precision of AAV lung gene therapy for SP-B deficiency using computationally derived lung-specific promoters.

Gene therapy·2026
Same journal

Recent advancements in improving cross-species applicability of bioengineered AAV capsids.

Gene therapy·2026
Same journal

Assessment of F/HN-pseudotyped lentiviral vector following intravenous delivery to mice.

Gene therapy·2026
Same journal

Applications of genome editing technologies in the treatment of human diseases.

Gene therapy·2026
Same journal

High resolution ES-DMA quantifies AAV capsid DNA content by electrical mobility to mass correlation.

Gene therapy·2026
Same journal

AAV8-mediated mouse/human PROC expression rescues thrombophilia in hereditary protein C-deficient mice.

Gene therapy·2026
See all related articles

Replication-competent retrovirus (RCR) unexpectedly emerged from a third-generation packaging cell line, GP + envAM12. This discovery has critical implications for human gene therapy safety protocols.

Area of Science:

  • Molecular Biology
  • Virology
  • Biotechnology

Background:

  • Third-generation retroviral packaging cell lines, like GP + envAM12, are designed to minimize recombination and prevent replication-competent retrovirus (RCR) formation.
  • These cell lines are crucial tools in gene therapy and retroviral vector production.

Purpose of the Study:

  • To report the unexpected discovery of RCR arising from routine use of the GP + envAM12 packaging cell line.
  • To investigate the characteristics and implications of this RCR for gene therapy applications.

Main Methods:

  • Transfection of GP + envAM12 cells with pBabeNeo plasmid to create a producer line.
  • Infection of indicator cell lines (B16, 1735-puro) with supernatant from the producer line.
  • Assessing gene transfer capabilities (neo, puro, lacZ) and receptor interference of the emergent RCR.

Related Experiment Videos

Main Results:

  • A replication-competent retrovirus (RCR) with an amphotropic envelope was detected in the GP + envAM12-pBabeNeo producer line.
  • The RCR successfully transferred genes (neo, puro, lacZ) to various cell lines, confirming its replication competence.
  • Minimal transfer to GP + envAM12 cells indicated receptor interference by the amphotropic envelope.

Conclusions:

  • The routine use of GP + envAM12 packaging cell lines can lead to the unintended generation of RCR.
  • This finding highlights potential safety risks associated with using such cell lines in human gene therapy.
  • Further safety assessments and modifications of packaging cell lines may be necessary for clinical applications.