Luciferase Tagging Cell Line Construction Service

Luciferase is a light-producing enzyme normally found in insect fireflies and luminous marine and terrestrial microorganisms. From transfection or fusion, expansion, screening, clone selection and titer assays, the Stable Cell Line Services offered by Seattle Genova are prepared to meet all of your custom cell line requirements. We have access to a wide array of technologies and a skilled team with experience with a range of cell line technologies. You will receive our top-notch customer service, which includes regular communication, quick turnaround times, and high success rates.

Applications of Luciferase Reporter Cell Lines

Cell-based assay systems using reporter enzymes are utilized widely to review promoters, interactions between promoters and transcription factors, signal transduction, and other cellular activities. Cell-based assays are also involved in drug screening both in vitro and in vivo. Of the reporter genes known to date, luciferases, enzymes that catalyze bioluminescence reactions, are utilized most continually because their sensitivity and linear response range are outstanding to those of normal reporters, including β-galactosidase, chloramphenicol acetyltransferase, β-glucuronidase, and green fluorescent protein.

◆Single Luciferase Reporter Assay

Luciferase is used as a reporter enzyme to estimate gene expression in prokaryotic or eukaryotic cells because the quantity of luciferase correlates with light intensity in the existence of excess luciferin and ATP. This system is adopted widely for basic biological studies, including those of gene expression, post-transcriptional modification, protein-protein interactions, and diagnostic and drug discovery applications because it is fitted for HTS. 

◆Dual Non-Secreted Luciferase Reporter Assay

Improvements in the transient transfection assay system have added luciferase as an internal control reporter (dual-reporter assay), thereby minimizing inherent experimental variability that can undermine experimental accurateness, such as variations in the number and viability of cells utilized, the efficiency of cell transfection and lysis, and so on. 

◆Dual Secreted Luciferase Reporter Assay

Secreted luciferases, CLuc, MetLuc, and GLuc, have been cloned from bright organisms such as the marine ostracod Cypridina noctiluca, and marine copepods Metridia longa and Gaussia princeps. As a reporter assay, the genes of CLuc, GLuc and MetLuc are used continually as commercial products. When cDNAs for these luciferases are transfected into mammalian cells, the expressed luciferases are emitted into the medium via the endoplasmic reticulum and the Golgi complex. The actions of the secreted luciferases can be quantified easily from the light intensity after the reaction with their substrates: CLuc reacts with Cypridina luciferin and GLuc and MetLuc react with coelenterazine. 

◆Multicolor Luciferase Reporter Assay

The dual luciferase assay system makes it feasible to conduct simultaneous monitoring of gene expression, intracellular detection of bioactive compounds, and HTS in vitro. However, for diagnostic and drug discovery purposes, both of which employ the analysis of a large number of samples, it is attractive to use an outstanding assay system whereby gene expression, protein-protein interaction, ligand binding, and post-transcriptional change can be regulated simultaneously in a one-step response in single-cell extracts. This system saves time and money, lessens the number of samples required, and promotes the interpretation of data. 

◆Real-Time Luciferase Reporter Assay

One benefit of using a luciferase reporter is that the luminescence can be calculated quantitatively and longitudinally with real-time resolution in vitro and in vivo when a bioluminescent substrate is conducted on cells. Among the possible luciferase-luciferin reactions, the firefly luciferase and firefly D-luciferin pair is utilized most widely, because the luminescence produced by the reaction can be quantified specifically, and has an incredibly low background, and because firefly D-luciferin is highly stable and easily permeates cells and tissues. Firefly luciferase and D-luciferin are sufficient for help as the reporter and substrate, respectively, for noninvasive measurements.

Cell line construction process 

◆Vector Construction 

Gene synthesis

Subcloning

Plasmid Preparation, QC sequencing 

◆Cell line development 

Host cell line analysis 

Viral/non-viral method

Stable cell pool generation 

◆Cell line Validation 

qPCR

FACS

Western blot

Immunofluorescence 

Stability test

◆Cell line Delivery 

Cell pool generation report

CoA report 

Highlights of Our services

◆Thousands cell delivery

◆Extensive proteins experience

◆PhD Expert Supports

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