Related Experiment Video
Updated: Aug 7, 2025
11:33
Microinjection of mRNA and Morpholino Antisense Oligonucleotides in Zebrafish Embryos.
Published on: May 7, 2009
36.9K
G-Quadruplexes Regulate miRNA Biogenesis in Live Zebrafish Embryos.
Tomás J Steeman1, Andrea M J Weiner1, Aldana P David1
1Instituto de Biología Molecular y Celular de Rosario (IBR), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad Nacional de Rosario (UNR), Ocampo y Esmeralda, Rosario S2000EZP, Argentina.
International Journal of Molecular Sciences
|March 11, 2023
Summary
RNA guanine quadruplexes (G4s) in pre-miRNA-150 regulate zebrafish development. Disrupting G4 formation enhances mature miRNA-150 levels and impacts gene expression, suggesting G4s control miRNA biogenesis.
Area of Science:
- Molecular Biology
- Developmental Biology
- Genetics
Background:
- RNA guanine quadruplexes (G4s) are regulatory structures influencing RNA metabolism.
- G4s in microRNA precursors (pre-miRNAs) may hinder their maturation by Dicer, affecting miRNA biogenesis.
- MicroRNAs (miRNAs) are crucial for embryonic development.
Purpose of the Study:
- Investigate the in vivo role of G4s in pre-miRNA-150 during zebrafish embryogenesis.
- Determine if G4 formation in pre-miR-150 impacts miRNA-150 biogenesis and subsequent gene expression.
- Explore the regulatory function of G4s in miRNA processing within a developmental context.
Main Methods:
- Computational analysis of zebrafish pre-miRNAs to identify putative G4 forming sequences (PQSs).
- In vitro folding and characterization of pre-miR-150 PQS.
- Microinjection of zebrafish embryos with modified pre-miR-150 (GTP vs. 7-deaza-GTP) and G4 stabilizer (pyridostatin).
- Analysis of mature miR-150 levels, target gene (myb) mRNA expression, and associated phenotypes.
Main Results:
- A conserved PQS in zebrafish pre-miR-150 was identified and shown to form G4 structures in vitro.
- Embryos injected with 7-deaza-GTP pre-miR-150 (unable to form G4s) exhibited increased mature miR-150 levels compared to GTP pre-miR-150.
- G4 formation in pre-miR-150 negatively regulated miR-150 biogenesis, leading to altered myb expression and developmental phenotypes.
- Stabilization of G4s with pyridostatin reversed these effects, rescuing gene expression and phenotypes.
Conclusions:
- The G4 structure in pre-miR-150 acts as a conserved regulatory element in vivo.
- G4 formation competes with the necessary stem-loop structure for miRNA biogenesis, thus controlling miRNA levels during zebrafish development.
- These findings highlight a novel mechanism of miRNA regulation mediated by G4 structures in non-coding RNAs.
