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

Esterase-triggered fluorescence of fluorogenic oligonucleotides

A Laurent1, F Debart, N Lamb

  • 1Laboratoire de Chimie Bio-organique, UMR 5625 CNRS-UM II, Université Montpellier II, France.

Bioconjugate Chemistry
|December 24, 1997
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

Comparison of surgical management and outcomes of acute right colic and sigmoid diverticulitis: a French national retrospective cohort study.

Techniques in coloproctology·2024
Same author

Risk factors for severe morbidity and definitive stoma after elective surgery for sigmoid diverticulitis: a multicenter national cohort study.

Techniques in coloproctology·2024
Same author

[Correlation between Objective structured clinical examinations (OSCE) and knowledge examination scores and analysis of factors associated with better OSCE success in fourth and fifth year medicine students at Dijon university hospital].

La Revue de medecine interne·2023
Same author

Prophylaxis is the new standard of care in patients with haemophilia.

South African medical journal = Suid-Afrikaanse tydskrif vir geneeskunde·2022
Same author

Hypertension in the South African public healthcare system: a cost-of-illness and burden of disease study.

BMJ open·2022
Same author

Effective Management Of Topical Nosocomial Aspergillus Spp. Infections In Three Severely Burned Patients.

Annals of burns and fire disasters·2021

This study developed a novel method to label antisense oligonucleotides with a fluorescent tag that is activated by cellular esterases. This allows for tracking the delivery and activation of these therapeutic molecules within cells.

Area of Science:

  • Oligonucleotide chemistry
  • Drug delivery systems
  • Molecular biology

Background:

  • Antisense oligonucleotides require intracellular delivery for therapeutic efficacy.
  • The prooligonucleotide approach utilizes cellular esterases for activation.
  • Efficient masking group removal is crucial for active oligonucleotide function.

Purpose of the Study:

  • To design and synthesize a method for labeling oligonucleotides with an esterase-activable fluorogenic tag.
  • To evaluate the ability of these labeled oligonucleotides to act as carboxyesterase substrates.
  • To visualize the intracellular delivery and activation of oligonucleotide conjugates.

Main Methods:

  • Synthesis of a biprotected carboxyfluorescein N-hydroxysuccinimide ester.

Related Experiment Videos

  • Conjugation of the tag to nuclease-resistant phosphorothioate and methylphosphonate oligodeoxynucleosides via an amino linker.
  • Assay of oligonucleotide conjugates for carboxyesterase substrate activity in various biological media.
  • Microinjection of phosphorothioate oligoconjugate into human fibroblasts for intracellular tracking.

    • Main Results:

    • Both oligonucleotide conjugates showed fluorescence upon incubation with esterases, serum, or cell extracts, indicating successful deprotection.
    • The phosphorothioate oligoconjugate demonstrated rapid fluorescence release in the cytoplasm after microinjection.
    • Translocation of the fluorescent oligomer into the nucleus was observed following cytoplasmic release.

    Conclusions:

    • The developed fluorogenic tag is effectively cleaved by cellular esterases, enabling the activation of prooligonucleotides.
    • This labeling method provides a valuable tool for monitoring oligonucleotide delivery and activation in real-time.
    • The findings support the use of esterase-activable prooligonucleotides for targeted intracellular delivery of antisense therapies.