RapidReporter®
RapidReporter®
• Vastly improved gene reporter vectors, reagents and associated technologies ideal for use in drug discovery, particularly HTS.
• Patented incorporation of both mRNA- and protein-destabilizing elements dramatically enhances both the speed and magnitude of response to drug effects.
• Combined with our patented synthetic luciferase, it provides the fastest and brightest reporter available.
• Provides a fast and inexpensive measure of gene expression or the activity of any upstream pathway.
• Numerous additional features provide greatly enhanced sensitivity and accuracy.
• Our Custom Vector service quickly gives you the reporter vectors you want, containing your targets of interest, with no fuss.
The RapidReporter® Concept
Reporter vectors link a gene’s regulatory element of interest (e.g. a promoter or a transcription factor binding site) to an easily measured reporter gene (e.g. a luciferase). Reporter levels are then used as a rapid measure of gene expression, or the activity of any signalling pathway upstream of the regulatory element.
However, molecules of reporter RNA and protein linger in a cell long after they were initially produced. The slow clearance rates of these reporter molecules substantially delays and dilutes the measurable response to stimulation or repression:
• In the case of repression, this is because the reporter mRNA continues to produce new, long-lived reporter protein long after transcription has stopped.
• In the case of stimulation, this is because the short, rapid increase in luciferase that occurs in response to whatever stimulation is being tested has proportionally little impact on the high steady-state levels of reporter already present.
In effect, much of the reporter protein being measured is derived from transcription that occurred before the treatment even began.
As a result, with standard reporter gene assays, transient or relatively minor effects are hidden, kinetic assays are inaccurate and the selective detection of active compounds is unreliable. Incorporating a protein-destabilizing element alone does not sufficiently solve this problem because new reporter protein continues to be produced as long as the pre-existing reporter mRNA remains intact.
Therefore, current commercially available gene reporter systems have not adequately addressed the confounding effects of both RNA and protein stability on assay results.
Until now.
Gene Stream’s patented RapidReporter® system utilises vectors that include both protein- AND mRNA- destabilizing elements, effectively eliminating the problem. No other reporter vectors feature this crucial innovation. (Examples)
We have developed a range of user-friendly, modular vectors. Each has been extensively optimised for maximum expression, sensitivity, response and accuracy.
Effect
RapidReporter® provides a faster and more pronounced response to drug effects that either activate or inhibit transcription (Examples). For example, upon treatment with a drug that inhibits transcription, the measurable signal of RapidReporter® drops to 50% of its initial levels about 12 times faster than the signal of a standard reporter. In a concentration response experiment, RapidReporter® was able to detect a biological response to an active compound at a concentration that was 50-fold lower than the minimum detectable concentration with a standard reporter.
An additional level of sensitivity of RapidReporter® is achieved by the profoundly optimised sequences that underlay each RapidReporter® construct. By enhancing reporter expression without affecting the kinetic behaviour, they provide the best of both worlds; high signal strength and superior responsiveness.
In drug-discovery applications, RapidReporter® provides a superior tool for target validation, lead identification/optimisation and efficacy studies. In particular, RapidReporter® vastly improves the selective detection of active compounds during HTS by substantially improving the measurable separation of active vs. inactive compounds. Benefits include faster assays, fewer spurious false positives, reduced impact of secondary effects and drug decomposition and most importantly, the confidence that you will no longer be throwing out valuable drug leads due to the inability of standard assays to detect their true activity.
Intracellular Synthetic Luciferase
The reporter gene of choice for the RapidReporter® system is our patented intracellular synthetic luciferase, derived via directed evolution of luciferases produced by the marine copepod Metridia family. This not only provides the fastest response of all known reporter genes but, when used with our luciferase assay kits, provides vastly superior signal strength compared to other reporter systems, including firefly or other coelenterazine luciferases like Renilla and Gaussia.
Whereas the use of destabilizing elements usually requires a considerable sacrifice in signal intensity, our sequence-optimised RapidReporter vectors used in combination with our luciferase assay kits, provide up to 140 times higher luminescence than that of even non-destabilized firefly luciferase.
Unlike the wild-type forms of secreted luciferases, which accumulate in the cell culture medium and thereby mask any changes in expression levels, our synthetic luciferase is not secreted, but is both intracellular and destabilized for maximum responsiveness.
Flexible licensing packages are available for commercial use of RapidReporter® technology within your own facilities.
*Voon, et al., Nucleic Acids Res, 2005.
Gene Stream Pty Ltd
Tel +61(0)408 903 609
