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

Targeted single-base correction by RNA-DNA oligonucleotides.

O Igoucheva1, K Yoon

  • 1Department of Dermatology and Cutaneous Biology, Jefferson Institute of Molecular Medicine, Jefferson Medical College, Philadelphia, PA 19107, USA.

Human Gene Therapy
|November 21, 2000
PubMed
Summary

RNA-DNA oligonucleotides (RDOs) can correct single-base mutations in mammalian cells, offering potential for treating genetic skin diseases. A new in vitro system enables rapid measurement of RDO gene correction efficiency.

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

White matter tracts that overlap with the thalamus and the putamen are protected against multiple sclerosis pathology.

Multiple sclerosis and related disorders·2021
Same author

Systemic CXCL10 is a predictive biomarker of vitiligo lesional skin infiltration, PUVA, NB-UVB and corticosteroid treatment response and outcome.

Archives of dermatological research·2021
Same author

Differential gene expression in laboratory strains of human head and body lice when challenged with Bartonella quintana, a pathogenic bacterium.

Insect molecular biology·2014
Same author

PPP1R1B-STARD3 chimeric fusion transcript in human gastric cancer promotes tumorigenesis through activation of PI3K/AKT signaling.

Oncogene·2013
Same author

Immunotargeting and eradication of orthotopic melanoma using a chemokine-enhanced DNA vaccine.

Gene therapy·2013
Same author

CHK2 kinase promotes pre-mRNA splicing via phosphorylating CDK11(p110).

Oncogene·2012

Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Single-base mutations cause genetic disorders, including skin diseases.
  • RNA-DNA oligonucleotides (RDOs) are a novel tool for gene correction.
  • Efficient and reproducible methods are needed to assess RDO efficacy.

Purpose of the Study:

  • To demonstrate RDO-mediated heritable gene correction in mammalian cells.
  • To establish a standardized in vitro system for measuring RDO gene correction.
  • To optimize RDO design and compare gene conversion frequencies across cell types.

Main Methods:

  • Development of an RNA-DNA oligonucleotide (RDO) for gene correction.
  • Demonstration of RDO-induced correction of a tyrosinase gene mutation in albino mouse melanocytes and skin.

Related Experiment Videos

  • Establishment of an in vitro nuclear extract system using a shuttle vector with a mutated E. coli beta-galactosidase gene.

    • Main Results:

    • RDOs successfully induced heritable correction of a point mutation at genomic, protein, and phenotypic levels.
    • The in vitro system accurately measured RDO-mediated gene correction in nuclear extracts.
    • The assay facilitated RDO design optimization and cell type-specific conversion frequency comparison.

    Conclusions:

    • RDOs show therapeutic potential for genetic skin diseases.
    • The developed in vitro system provides a rapid and reproducible method for assessing RDO efficiency.
    • Further mechanistic studies using this system will advance RDO technology for site-specific gene correction.