3'UTR Virus
Product DetailsApplicationCase StudyFAQ
Product Details
miRNAs function by binding to the 3' untranslated region (3'-UTR)of target mRNAs,leading to mRNA degradation or translational repression.With over 2600 miRNAs identified in humans,a single miRNA has the potential to regulate hundreds of genes,indicating their widespread influence on gene expression.
Creative Biogene offers a comprehensive 3'UTR lentiviral vector library targeting all human genes, featuring a choice
of GFP or luciferase reporter genes. These constructs incorporate 3'-UTR sequences obtained from public gene sequence databases,cloned downstream of a reporter gene in a lentiviral vector under the control of the CMV promoter.Creative Biogene's 3'UTR lentiviral vector library provides researchers with a powerful platform for exploring miRNA-mediated gene regulation and its implications in diverse biological contexts.
Key Features of Our 3'UTR Virus
- Creative Biogene's Solution:Comprehensive coverage of all human genes.
- Flexible Reporter Options:Choice between GFP or luciferase reporter genes.
- Reliable Source:Incorporates 3'-UTR sequences from reputable public databases.
- Trusted Platform: Empowers researchers to explore miRNA-mediated gene regulation effectively.
3'UTR Virus List
Application
Our 3'UTR Virus platform enables efficient investigation of miRNA-mediated gene regulation, facilitating
comprehensive studies on cellular processes and disease mechanisms. Leveraging our advanced technology and extensive
library targeting all human genes, researchers can explore miRNA binding requirements with ease and accuracy:
- miRNA Target Validation: Co-transfect the 3'UTR reporter vector with miRNA expression vectors or mimics.Reduced expression of the reporter gene indicates validated interaction between the 3'UTR and the
miRNA.
- Endogenous miRNA Expression Assay: Transfect the 3'UTR reporter vector into the cell line of interest to assay for endogenous miRNA expression.Co-transfection with miRNA inhibitors should result in reduced or abolished interactions between the miRNA and the UTR of the Reporter Vector.
- Investigation of miRNA Binding Requirements:Use 3'UTR mutant reporters to investigate miRNA
binding requirements. Mutate specific loci within the 3'UTR and observe changes in reporter gene expression to determine the importance of each locus for miRNA binding.
- Stable Expression of 3'UTR Reporter Construct: Establish 3'UTR reporter cell lines for stable expression of the 3'UTR reporter construct derived from HEK293 cells. This enables continuous monitoring of
miRNA-mediated regulation in a stable cellular environment.
Case Study
Case Study 1
The role of cellular senescence as a tumor suppression mechanism highlights the importance of investigating gene
expression regulation during senescence, providing insights into cancer therapy. Researchers aimed to elucidate the
role of the 3'UTR Virus vector in investigating the regulatory mechanisms of gene expression during cellular senescence,with implications for cancer therapy.By studying alternative polyadenylation(APA)and its impact on the HN1 gene,they found that APA of HN1 correlated with cancer and senescence,influencing its stability and protein production.Manipulating HN1 expression induced senescence-associated phenotypes,and patients with higher HN1 expression exhibited lower survival rates in various carcinomas.Notably,modulating the splicing factor HNRNPA1 affected HN1 APA and senescence-associated phenotypes,indicating the HNRNPA1-HN1 axis as a potential target for cancer treatment.This study underscores the importance of understanding gene regulation in senescence and its implications for cancer therapy.
Figure 1.Researchers investigated the downstream effects of HN1 transcripts with shorter(HN1-S)and longer(HN1-L)3' UTRs on gene expression, respectively, using 3'UTR Virus vectors.(Jia Q,et al.,2019)
Case Study 2
In colorectal cancer(CRC),PLAGL2 or POFUT1 promote tumorigenesis and exhibit significant positive correlations,yet the mechanism driving their co-expression and functional implications remain unclear.Researchers investigated the co-expression mechanism and functional implications of PLAGL2 and POFUT1 in colorectal cancer(CRC).Clinical data and TCGA dataset analysis confirmed their significant positive correlation(r=0.91,p<.0001).Employing luciferase reporter assays,bidirectional promoter vectors,and 3'UTR expression, they elucidated an
evolutionarily conserved bidirectional promoter responsible for their co-expression. Furthermore, in vitro and
in vivo assays demonstrated their synergistic oncogenic effects in CRC, particularly in maintaining colorectal
cancer stem cell (CSC) stemness via the Wnt and Notch pathways. Mutational analysis of shared transcription
factor binding sites confirmed the regulatory role of the bidirectional promoter. This study uncovers the
oncogenic potential of the PLAGL2-POFUT1 bidirectional promoter pair, offering novel therapeutic targets for CRC
intervention.
Figure 2. Lentiviral vectors carrying PLAGL2, POFUT1, and
various cDNA constructs, including PLAGL2–3'UTR1 and PLAGL2–3'UTR2, were utilized, along with lentiviral
shRNA targeting human PLAGL2 and POFUT1. (Li D, et al., 2019)
FAQ
Q: How do these 3'UTR Virus products assist in validating the interaction between miRNA and target genes?
A:Our 3'UTR Virus platform enables researchers to effectively validate the interaction between
miRNA and target genes by observing the reduction in reporter gene expression to confirm whether
such interaction occurs.
Q: How are these products utilized to analyze the expression of endogenous miRNA?
A: By transfecting the 3'UTR reporter vector into the cell line of interest,along with miRNA inhibitors,one can observe whether the interaction between miRNA and the UTR of the Reporter Vector is reduced or abolished,facilitating the analysis of endogenous miRNA expression.
Q:How do these products aid in studying the binding requirements of miRNA?
A:By employing 3'UTR mutant reporters, researchers can determine the significance of a locus for
miRNA binding. Mutating the locus and observing whether reporter gene expression is affected
allows for the exploration of miRNA binding requirements.
Q: How are 3'UTR reporter cell lines utilized for stable expression of the 3'UTR reporter
construct?
A: Our products offer cell lines for stable expression of the 3'UTR reporter construct from HEK293 cells,providing researchers with a more stable and reliable expression system to facilitate their research endeavors.
* For research use only. Not intended for any clinical use.