GeneCopoeia: Gluc-ON™ Promoter Reporter Clones

By: Genecopoeia  11-11-2011

Using a secreted and robust Gaussia Luciferase (GLuc) as the reporter, GeneCopoeia GLuc-ON™ promoter clones are designed to detect the real-time activities of over 20,000 human promoters using live cell assays.

Each transfection-ready promoter clone contains a 1.2-1.5 kb insert, corresponding to the 5'-flanking sequence located approximately 1.5 kb upstream and up to 200 bp downstream of the transcription start site (TSS) of a specific human gene. This insert is placed upstream of the GLuc reporter gene. Since the putative cis-acting enhancer elements are expected to exist in the cloned promoter region, the luciferase activity observed during the reporter assay closely resembles the actual promoter regulation of these genes within human cells.

Figure 1. How GLuc-ON promoter clones work.

Advantages

    Live cell assays
  • Naturally secreted GLuc reporter
  • No lysis of the cells is necessary
  • Save samples, reduce variations, and simplify experiments for applications such as pulse chase analysis,etc.
    Real-time study
  • Data is generated quickly
  • Closely resembles real-time activities
    Dual secreted reporter system
  • Secreted GLuc and SEAP
  • Enables transfection-normalization for accurate across-sample comparison
    High-throughput compatible
  • Group or pathway study compatible
  • High sample number compatible
    High sensitivity
  • GLuc is 1000-fold more sensitive than firefly or Renilla luciferase
    Convenience
  • All promoter clones are transfection-ready

Gaussia luciferase

GLuc-ON promoter clones use a modified GLuc (mGLuc) as the reporter gene, which generates a highly stable signal and overcomes the quick signal decay commonly observed with humanized wild type GLuc (wtGLuc).

Figure 2. Signal stability of mGLuc (blue) and wtGLuc (red). Left: assay buffer with a stabilizer; Right: regular assay buffer (Secrete-Pair™ dual luminescence assay kit)

Dual-reporter system

Dual-reporter vectors are available for the GLuc-ON promoter clones. The secondary reporter, secreted Alkaline Phosphatase (SEAP), serves as an internal control. The dual-reporter system enables transfection normalization for accurate cross-sample comparison.

Figure 3. Normalized promoter activities in H1B1B and HEK293T cells. Dual-reporter promoter clones or controls were transfected into two cell lines in duplicates. Samples were analyzed 24 hrs (HEK293T) and 48 hrs (H1B1B) after transfection. NEG (containing non-promoter sequence) and EMPTY (no promoter in the vector) are negative controls.

Vector Reporter gene Tracking gene Selection marker
Gaussia luciferase (GLuc) N/A* N/A
Gaussia luciferase (GLuc) N/A* Puromycin
Gaussia luciferase (GLuc) Secreted alkaline phosphatase (SEAP) N/A
Gaussia luciferase (GLuc) Secreted alkaline phosphatase (SEAP) Puromycin
eGFP N/A* N/A
eGFP N/A* Puromycin

* A separate vector is available for SEAP expression.

Custom promoter clone services

Want to compare various length or regions of your promoter? You can custom-order them.
Email:
Call:866-360-9531 or 301-762-0888

Control clones and SEAP expression clone


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