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Updated: Dec 11, 2025
Genome-wide Screen for miRNA Targets Using the MISSION Target ID Library
Published on: April 6, 2012
[Analysis of miRNA-326's action on its target gene BCL-XL]
Mingming Qiao1, Xia Gai, Hui Ye
1Blood Center of Shandong Province, Jinan, Shandong 250014, China. happymike110@126.com.
Objective:
To analyze the action of miRNA-326 on its target gene BCL-XL and the molecular mechanism of platelet apoptosis regulated by miRNAs.
Methods:
Dual-luciferase vectors containing respectively the wild-type and mutant 3'-untranslated region (3'UTR) fragments of the BCL-XL gene were constructed with firefly and renilla luciferases and transfected into 293T cells. Relative fluorescence intensities of the transfected cells were measured.
Results:
Dual-luciferase reporter gene vectors for PsiCHECK- BCL-XL -3'UTR-WT (wild-type) and PsiCHECK- BCL-XL -3' UTR-MT (variant) were respectively constructed. Relative fluorescence intensities of the 293T cells co-transfected by miRNA-326 and PsiCHECK- BCL-XL -3'UTR-WT plasmid were significantly lower compared with the control group (co-transfected by a miRNA-326 negative sequence and PsiCHECK- BCL-XL -3' UTR-WT plasmid) ( P = 0.034). The relative fluorescence intensity was also significantly reduced in cells co-transfected by miRNA-326 and PsiCHECK- BCL-XL -3' UTR-WT plasmid compared with the mutant control group co-transfected by miRNA-326 and PsiCHECK- BCL-XL -3'UTR-MT plasmid (P = 0.022).
Conclusion:
miRNA-326 may participate in the regulation of platelet apoptosis by acting on the 3'-UTR of the BCL-XL gene.
Insights
MicroRNA-326 (miRNA-326) targets the BCL-XL gene, influencing platelet apoptosis. This study reveals miRNA-326
Area of Science:
- Molecular Biology
- Cell Biology
- Hematology
Background:
- MicroRNAs (miRNAs) are key regulators of gene expression.
- Platelet apoptosis is a critical process in hemostasis and thrombosis.
- BCL-XL is an anti-apoptotic protein with a role in platelet survival.
Purpose of the Study:
- To investigate the interaction between miRNA-326 and its target gene BCL-XL.
- To elucidate the role of miRNAs in regulating platelet apoptosis.
Main Methods:
- Construction of dual-luciferase reporter vectors with wild-type and mutant 3'-untranslated regions (3'UTR) of BCL-XL.
- Transfection of these vectors into 293T cells.
- Measurement of relative fluorescence intensities to assess miRNA activity.
Main Results:
- Dual-luciferase reporter assays confirmed that miRNA-326 directly targets the 3'-UTR of BCL-XL.
- Co-transfection with miRNA-326 significantly reduced luciferase activity in cells containing the wild-type BCL-XL 3'-UTR.
- This reduction was more pronounced compared to control groups with a negative miRNA sequence or a mutated BCL-XL 3'-UTR.
Conclusions:
- miRNA-326 negatively regulates BCL-XL expression.
- miRNA-326 is implicated in the molecular mechanism of platelet apoptosis via targeting BCL-XL.
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