Studies of protein expression, interactions, and structure, often use reporter systems to introduce and select for gene targets in cells. Reporter genes confer drug resistance, bioluminescence or fluorescence properties into the cells they are introduced. Typical reporter studies link reporter genes directly to a promoter region of interest, the function of which can be monitored by the reporter activity. Protein fusion tagging is used to detect subcellular localization, degradation, protein-protein interactions, etc. Typical fusion tags are fluorescent proteins (e.g eGFP) or small protein epitopes (e.g FLAG, Myc HA) which can be detected by fluorescence FACS or western blots. New generations of reporter gene systems expand the range of applications and enhance experimental possibilities.
- Luciferase reporter genes with the enhanced properties of greater stability, robust secretory signal, smaller size, high sensitivity and no dependence on ATP are applicable for reporter gene studies, siRNA potency screening, live cell monitoring and BRET applications in vivo.
- Enhanced GFP and GFP color derivatives are useful for imaging mammalian cells, single fluorescent protein-based biosensors, FRET and producing transgenic model organisms. However GFP can be toxic to cells at high expression levels.
- Multimodality fusion reporter technologies employ vectors containing multiple reporter genes. These systems enable optimal analysis by multiple methods, such as fluorescence for single cell imaging, bioluminescence for small animal imaging, or short-lived radioactive tracers in living subjects for PET.
- Split reporter systems enable protein-protein interaction assays by cloning target genes into separate vectors, each linked to complementary portions of a reporter, such as luciferases or GFP.
Reporter Systems includes these areas of focus:
Protocols for Reporter Systems
- Cellular Labeling (E9100)
- Cellular Labeling (E9120)
- Cellular Labeling (E9230)
- Cloning of SNAP-tag Fusions in pSNAPf (N9183)
- Expression of SNAPf Fusions (N9183)
- Instructions for Cellular Labeling (E9200)
- Labeling of Proteins in Solution (E9230)
- Labeling of Proteins in vitro (E9100)
- Labeling Proteins in vitro (E9120)
- Labeling Proteins in vitro (E9200)
- Use of CLIP-Cell Block with CLIP-Cell Substrates (E9230)
- Use of SNAP-Cell Block with SNAP-Cell Substrates (E9100)
- Western Analysis (E8023)
Application Notes Reporter Systems
Gaussia and Cypridina Luciferases - Ultrasensitive secreted reporters and their use in dual assays
Other Tools & Resources
NEB vs. Other Commercially Available Reporter Systems
|Does not require and
not affected by ATP
Comparison of the features of commercially available reporter systems highlights the advantages of using Gaussia or Cypridina Luciferase from NEB.