Report Gene Vector Building Service

Lifeasible can provide cutting-edge, comprehensive, and rigorous reporter gene vector design and vector construction services for gene function researchers, including untranslated region (UTR) reporter gene plasmid, promoter luciferase reporter gene plasmid, single-nucleotide polymorphism (SNP), and other reporter genes. Among them, the luciferase reporter gene system is a reporter system that uses luciferin as a substrate to detect luciferase activity. Firefly luciferase and renilla luciferase are two commonly used reporter genes.

Service flow (luciferase reporter gene vector building as an example)

Sample requirements

• Detailed information about transcription factors, target genes, or microRNAs is required. If there are gene templates or constructed expression vectors, they can be provided directly.
• Experimental cells. If no special requirement exists, 293T cells will be used by default.

Delivery

One copy of the experimental procedure and a complete report, including experimental raw data, pictures, analysis results, etc.

Advantages

• Fast expression time, target gene expression can be observed about 24 hours after cell introduction.
• Our overexpression vector library is abundant, and the overexpression markers are abundant.
• Fast building process and low cost.

Applications

• Verification of miRNA interactions with target genes mRNA targeting interactions

The predicted target sequence of the target gene to be tested is inserted into the 3'-UTR region of F-Luc in the reporter gene vector and then co-transfected into the miRNA mimics. If luciferase activity is decreased, it suggests that the sequence to be sequenced is the target sequence of the miRNA.

• Promoter/enhancer structure analysis

The promoter region sequence (usually around 2 k) is segmentally truncated or mutated to a specific site. The promoter sequences are then constructed into luciferase reporter vectors to replace the promoter sequences, and their promoter activities are detected.

• Verification of specific transcription factors interacting with their regulatory sequences

The predicted promoter sequences interacting with the transcription factors were inserted into the reporter vectors, and the transcription factors were co-transformed and overexpressed in the experimental cells. Overexpression of the transcription factor can be analyzed to see if it increases luciferase activity.

• Verification of signaling pathway activation

A pathway-related downstream response element is constructed into the promoter region of the reporter gene vector, and luciferase activity is increased if the pathway is activated.

Lifeasible can construct specific fragments into reporter gene vectors according to the customer's experimental needs and the target genes to be studied, which can be used to research gene expression regulation, signaling pathways, SNPs, and other related studies. Please feel free to contact us to start your research.