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

Antisense principle or ribozyme action?

G Sczakiel1, R S Goody

  • 1Deutsches Krebsforschungszentrum, Heidelberg, Germany.

Biological Chemistry Hoppe-Seyler
|November 1, 1994
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

Time-Resolved Structural Studies on Insect Flight Muscle after Photolysis of Caged-ATP.

Biophysical journal·2009
Same author

Phosphorothioate-stimulated cellular uptake of siRNA: a cell culture model for mechanistic studies.

Current pharmaceutical design·2008
Same author

[Breast cancer diagnostics based on extracellular DNA and RNA circulating in blood].

Biomeditsinskaia khimiia·2008
Same author

The structural and mechanistic basis for recycling of Rab proteins between membrane compartments.

Cellular and molecular life sciences : CMLS·2005
Same author

[Ki-67 antisense therapy in murine renal cell carcinoma models].

Aktuelle Urologie·2005
Same author

Spontaneous uptake of biologically active recombinant DNA by mammalian cells via a selected DNA segment.

Gene therapy·2004

Incorporating hammerhead ribozyme catalytic domains into antisense RNA enhances cellular inhibition. In vitro kinetics do not fully explain in vivo ribozyme activity, suggesting additional factors are involved.

Area of Science:

  • Molecular Biology
  • RNA Therapeutics
  • Biochemistry

Background:

  • Antisense RNA (AS RNA) is a tool for gene silencing.
  • Hammerhead ribozymes are catalytic RNA molecules with self-cleaving activity.
  • Understanding ribozyme mechanisms in vivo is crucial for therapeutic applications.

Purpose of the Study:

  • To investigate the impact of hammerhead ribozyme catalytic domains on antisense-mediated gene inhibition in living cells.
  • To evaluate the compatibility of in vitro kinetic models with observed in vivo ribozyme function.

Main Methods:

  • Incorporation of hammerhead ribozyme catalytic domains into long antisense RNA molecules.
  • Assessing antisense-mediated inhibition in cellular systems.
  • Kinetic analysis of RNA duplex formation and target RNA cleavage in vitro.

Related Experiment Videos

Main Results:

  • Hammerhead ribozyme catalytic domains significantly enhanced antisense-mediated inhibition in living cells.
  • In vitro kinetic parameters for duplex formation and cleavage were insufficient to explain the in vivo effects.
  • Discrepancies suggest the need for additional assumptions to model in vivo ribozyme action.

Conclusions:

  • Hammerhead ribozymes are effective enhancers of antisense RNA activity in cellular environments.
  • Current kinetic models require refinement to accurately predict ribozyme behavior in vivo.
  • Further research is needed to elucidate the in vivo mechanisms governing ribozyme function.