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

Substrate requirements for let-7 function in the developing zebrafish embryo.

Wigard P Kloosterman1, Erno Wienholds, René F Ketting

  • 1The Hubrecht Laboratory, Centre for Biomedical Genetics, 3584 CT Utrecht, The Netherlands.

Nucleic Acids Research
|December 9, 2004
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

Crosstalk between and developmental dynamics of Caenorhabditis elegans Argonaute proteins.

Genetics·2026
Same author

Tdrd6a and Tdrd6b are required together for germ plasm formation.

Development (Cambridge, England)·2026
Same author

SLBP-independent control of maternal histone mRNA.

bioRxiv : the preprint server for biology·2026
Same author

Multi-scale simulations of MUT-16 scaffold protein phase separation and client recognition.

Biophysical journal·2025
Same author

Meioc-Piwil1 complexes regulate rRNA transcription for differentiation of spermatogonial stem cells.

eLife·2025
Same author

A genetic framework for RNAi inheritance in Caenorhabditis elegans.

EMBO reports·2025
Same journal

VeloRM: disentangling pre- and post-splicing RNA modification dynamics at single-cell resolution.

Nucleic acids research·2026
Same journal

Accessibility of telomeric overhangs to stabilizing small-molecule ligands.

Nucleic acids research·2026
Same journal

Multivalent interactions mediate SNAIL transcription factor stimulation of the nucleosome deacetylase activity of the CoREST complex.

Nucleic acids research·2026
Same journal

Genome-wide mapping of DNA G-quadruplexes in Trypanosoma brucei chromatin reveals enrichment in coding regions and transcription start sites.

Nucleic acids research·2026
Same journal

Epigenetic landscape of synthetic human centromere core regions: CENP-A assembly and euchromatic modifications interdependently antagonize heterochromatin accumulation.

Nucleic acids research·2026
Same journal

MAPK/ERK signaling regulates H3K9me3 heterochromatin reorganization to confer mesendoderm developmental competence.

Nucleic acids research·2026
See all related articles

The let-7 microRNA (miRNA) silences gene expression in zebrafish by binding to target sites. Its 5' region is crucial for function, and multiple miRNAs can cooperate to enhance silencing.

Area of Science:

  • Molecular Biology
  • Developmental Biology
  • Genetics

Background:

  • MicroRNAs (miRNAs) regulate gene expression post-transcriptionally by binding to messenger RNAs (mRNAs).
  • The let-7 miRNA family is highly conserved across species and plays a vital role in development.
  • Understanding miRNA target recognition is crucial for deciphering gene regulatory networks.

Purpose of the Study:

  • To investigate the substrate requirements for let-7 microRNA function in a vertebrate in vivo system.
  • To identify the critical regions of let-7 miRNA necessary for its gene silencing activity.
  • To explore cooperative interactions between different miRNAs in gene regulation.

Main Methods:

  • Utilized zebrafish embryos as a vertebrate model system for in vivo studies.

Related Experiment Videos

  • Injected double-stranded let-7 miRNA into zebrafish and frog zygotes to observe phenotypic effects.
  • Employed reporter assays with green fluorescent protein (GFP) fused to target 3'-untranslated regions (UTRs) to assess silencing.
  • Analyzed point mutant derivatives of let-7 to determine functional domains.

    • Main Results:

    • The antisense strand of let-7 mediates biological activity and causes specific phenotypic defects.
    • let-7 effectively silences mRNA containing its target sites in the 3'-UTR of a lin-41 ortholog.
    • Both let-7 target sites in lin-41 3'-UTR are essential and sufficient for silencing.
    • Cooperative silencing was observed when let-7 and mir221 acted together on a modified target construct.
    • let-7 target sites are functional when located in coding sequences or 5'-UTRs.
    • Analysis of let-7 point mutants revealed that the 5' region is critical for its function.

    Conclusions:

    • The 5' region of let-7 is essential for its gene silencing activity.
    • Multiple miRNAs can cooperate to achieve robust gene silencing.
    • Zebrafish embryos provide a powerful system for studying miRNA function and target recognition in vivo.