Product Details

Creative Biogene offers genome-wide human, mouse, and rat miRNA 3' untranslated region (3'UTR) target clones in mammalian expression vectors. These clones contain the 3'UTR sequence downstream of a reporter gene driven by the SV40 promoter. They are designed to study miRNA-mediated post-transcriptional regulation by quantifying differential luciferase activity. These clones can be used for miRNA target identification, functional verification of predicted targets, and studying the regulatory effects of miRNAs on target genes.

Advantages of Our miRNA 3'UTR Clones

• Genome-wide Coverage: Our miRNA 3'UTR target clones cover the entire human, mouse, and rat genomes, providing comprehensive resources for miRNA target identification and functional studies
• High-Quality Sequences: The 3'UTR sequences used in our clones are obtained from public domain gene sequence databases, ensuring accuracy and reliability in experimental results.
• Mammalian Expression Vector: The clones are constructed in mammalian expression vectors, allowing for efficient expression and functional analysis in mammalian cells.
• Versatile Applications: These clones can be utilized for various applications, including miRNA target identification, functional validation, and investigation of miRNA-mediated gene regulation.
• Quantifiable Results: The differential luciferase activity generated by the interaction between the 3'UTR sequences and miRNAs can be quantified using a colorimetric assay, providing precise and measurable data for analysis.

miRNA 3'UTR Clone List

Application

MicroRNAs (miRNAs) are short regulatory RNAs, approximately 20 to 22 nucleotides in length, found in both plants and animals. In humans, more than 400 miRNAs are encoded by approximately 4% of the genome, and they are estimated to regulate at least 30% of human genes. These miRNAs play crucial roles in various biological processes by binding to complementary sequences in the 3'UTR of target mRNAs. In animals, most miRNAs form imperfect duplexes with sequences located in the 3'UTR region of target mRNAs. In light of these functions, our products offer valuable tools for researchers engaged in understanding miRNA-mediated gene regulation and its implications in various biological contexts. Specifically, our products can facilitate the following research endeavors:

• miRNA Target Validation: These miRNA 3'UTR clones validate gene targets for miRNAs. Researchers transfect or transduce these clones into cells, observing miRNA's regulatory effect by examining its interaction with the target gene's 3'UTR.
• Gene Expression Regulation Studies: These clones investigate the 3'UTR's role in gene expression regulation. Analyzing miRNA-target gene interactions deepens understanding of post-transcriptional regulation mechanisms.
• Signal Transduction Pathway Research: These clones study gene regulation in signal transduction pathways, revealing miRNA's role in modulating cellular signaling.
• Drug Target Screening: Studying these miRNA 3'UTR clones in cells identifies potential drug targets, aiding in developing novel disease therapies.
• Exploration of Disease Mechanisms: Applying these clones in models explores disease mechanisms, and understanding disease progression by investigating miRNA's regulatory effect on target genes.

Case Study

Case Study 1

Acute myeloid leukemia (AML) is a highly aggressive cancer that affects individuals of all ages worldwide. The researchers investigated the potential therapeutic effects of Ginsenoside Rh2 (GRh2) on pediatric leukemia. They conducted experiments using mouse models and leukemia cell lines to evaluate the impact of GRh2 on leukemia cell survival and apoptosis. Their findings demonstrated that GRh2 significantly prolonged the survival of mice with pediatric leukemia and decreased the viability of leukemia cells in vitro by inducing apoptosis. Further analysis revealed that GRh2 reduced the levels of the anti-apoptotic protein Bcl-2 in leukemia cells. Interestingly, they discovered that GRh2 may exert its effects by inducing the expression of microRNA-21 (miR-21), which in turn suppressed the translation of Bcl-2 mRNA by binding to its 3'-UTR.

Figure 1. miR-21 significantly reduced the luciferase activity of 3'-UTR reporter for Bcl-2, indicating its role in suppressing Bcl-2 translation. Thus, GRh2-induced apoptosis in leukemia cells may occur through miR-21-mediated suppression of Bcl-2. The Bcl-2 3'-UTR reporter plasmid (pRL-Bcl-2) was purchased from Creative Biogene. (Wang X, et al., 2015)

Case Study 2

Endothelial cell injury and death are crucial in atherosclerosis, with autophagy's protective role unclear; miR-30, an autophagy suppressor, may play a role in atherosclerosis development. Researchers aimed to elucidate the molecular regulation of endothelial cell autophagy in atherosclerosis by investigating the role of miR-30 and its interaction with ATG6 mRNA. Using a mouse model for atherosclerosis, they found that high-fat diet (HFD) led to decreased levels of endothelial cell autophagy and ATG6 protein, but not mRNA. Bioinformatics analyses and luciferase reporter assays confirmed the binding of miR-30 to the 3'-UTR of ATG6 mRNA, suggesting a regulatory mechanism at the post-transcriptional level. In vitro experiments on oxidized low-density lipoprotein (ox-LDL)-treated human aortic endothelial cells (HAECs) further supported these findings. The upregulation of miR-30 by HFD was associated with impaired endothelial cell autophagy, potentially contributing to the development of atherosclerosis.

Figure 2. miR-30 was identified as targeting the 3'-UTR of ATG6 mRNA, with its levels significantly increased after HFD treatment. By modifying miR-30 levels in HAECs and using an ATG6 3'-UTR reporter plasmid, it was demonstrated that ATG6 translation is suppressed by miR-30, potentially contributing to impaired endothelial cell autophagy in atherosclerosis. The ATG6 3'-UTR reporter plasmid (pRL-ATG6) was purchased from Creative Biogene. (Zhang T, et al., 2015)

Case Study 3

The identification of microRNAs involved in regulating bladder cancer metastases holds promise for uncovering novel therapeutic targets in BC treatment. Researchers aimed to investigate the role of miR-138 in regulating bladder cancer (BC) cell invasion and metastases. They analyzed miR-138 and ZEB2 levels in BC specimens, finding miR-138 significantly decreased while ZEB2 increased compared to normal bladder tissue, with metastatic BC showing lower miR-138 levels. Through bioinformatics analyses and luciferase reporter assays with the ATG6 3'-UTR reporter plasmid, they confirmed miR-138 targeting of ZEB2 mRNA's 3'-UTR to inhibit translation. Additionally, miR-138 overexpression suppressed ZEB2-mediated invasion and metastases, while depletion increased them in BC cells. These findings suggest that miR-138 suppression in BC cells may promote ZEB2-mediated cancer invasion and metastases, highlighting miR-138 as a potential therapeutic target for preventing BC metastases.

Figure 3. Through bioinformatics analyses, researchers identified miR-138 binding sites on the 3'-UTR of ZEB2 mRNA, spanning from the 766th to the 772nd base site. Target plasmids for 3'-UTR of ZEB2 mRNA clone or 3'-UTR of ZEB2 mRNA with a mutant at miR-138 binding sites wre purchased from Creative Biogene. (Sun DK, et al., 2015)

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