LightSwitch™ Validated Pathway Collections

pathway-specific, validated luciferase reporter constructs

LightSwitch™ Validated Pathway Collections contain promoter reporter constructs selected from the 18,000-member LightSwitch Promoter Reporter GoClone® Collection. Each collection was created using pathway-specific induction experiments, so all members have been experimentally validated to be well suited to study a specific pathway of interest, such as inflammation, hypoxia, DNA damage, etc.

Heat Map

Heat Map of Representative Reporter Constructs versus Cell treatments.

To prepare these validated constructs, promoter sequences for each collection were chosen based on motif analysis and published information. In-house luciferase assays were then performed, and only those promoter reporter constructs that showed significant pathway induction response (activation or repression) were included in the collections. More information for each transcription factor is listed below.

Validated Pathway Collections

Transcription Factors

NFΚB is a ubiquitous transcription factor that regulates the genes that mediate cell cycle, apoptosis, intercellular communication, immune response and development, memory formation, and tumorigenesis. Additionally, various signals such as intracellular stress, cytokines, UV irradiation, free radicals, and infection induce gene expression through the NFκB family. Understanding the NF-kb pathway may aid in identifying preventive and therapeutic measures for diseases, such as chronic inflammation and cancer. More >

Pathway: Inflammation

Induction Treatment: TNF-Α

The CREB (cAMP responsive element binding protein) signaling pathway regulates a variety of biological processes including cell survival and proliferation, glucose regulation, neuronal differentiation, and long-term memory formation. The richness of CREB signaling illustrates its significance as a key regulator of the expression of a multitude of physiologically important genes.
More >

Pathway: cyclic-AMP

Induction Treatment: Forskolin, PMA

Hypoxia, low oxygen concentration in cells, is a characteristic of various physiological processes such as the regulation of cell differentiation, blood vessel formation, and wound healing. Hypoxia is also widely associated with diseases such as anemia, diabetes, kidney disease, cardiovascular disease, hypertension, sickle cell disease, and many cancers. Cells under hypoxic stress activate a signaling cascade of hypoxia-responsive genes to aid the cell into either adapting to the low oxygen conditions or undergoing apoptosis. This response to restore cellular homeostasis is primarily initiated and regulated by the family of hypoxia inducible transcription factors (HIF-1α). More >

Pathway: Hypoxia

Induction Treatment: 1% Oxygen, DFO

STAT1 belongs to the family of Signal Transducers and Activator of Transcription (STAT), which regulates cell growth, differentiation, survival and apoptosis. STAT1 also plays a role in immune response, and it mediates interferon signaling. Understanding the mechanisms of how STAT1 regulates various gene pathways gives further insight into the roles of STAT1 in normal development, pathology and targeted drug development. More >

Pathway: Interferon response

Induction Treatment: Interferon-α, interferon-γ

The role of p53 in tumor suppression is well characterized. A broad range of stressors such as hypoxia, DNA damage, and viral infection stabilize and activate p53, inducing a large cascade of genes that play significant roles in cell cycle arrest, DNA repair and apoptosis. More >

Pathway: DNA damage, apoptosis

Induction Treatment: Nutilin

Heat shock proteins (HSPs) are also well-characterized for their roles as chaperones for protein synthesis, assembly, secretion and degradation. Specific HSPs have been implicated as central players in apoptosis, tumor survival, and immune response, and an understanding of the regulation of the heat shock response is critical. More >

Pathway: Heat shock

Induction Treatment: 43ºC

The sterol regulatory element binding proteins (SREBPs) are transcription factors that regulate lipogenesis. The SREBP isoforms, SREBP-1a, SREBP-1c, and SREBP-2, specifically control the expression of genes involved in cholesterol, fatty acid, triglyceride and phospholipid synthesis. SREBP-1c has also been implicated in glucose uptake regulation. Dysregulation of SREBP and the SREBP-regulated pathways is clinically relevant in disorders such as diabetes mellitus, insulin resistance, non-alcoholic fatty liver disease (NAFLD) and obesity. Understanding the mechanisms of how SREBPs are coordinated with other physiological pathways can provide further insight into the roles of SREBP in normal development, and can lead to successful strategies for targeting SREBP pathological research and drug therapies. More >

Pathway: Cholesterol biosynthesis

Induction Treatment: Statins, Cholesterol

Serum response factor (c-Fos serum response element-binding transcription factor, or SRF) is a transcription factor that regulates the activity of various genes involved in cell cycle, apoptosis, embryogenesis, cell growth, neuronal transmission, and cell differentiation. Improper regulation of SRF pathways has been implicated in various disorders including ulcers, psoriasis, and cardiomyopathy. Understanding the mechanisms of how SRF is coordinated with other physiological pathways can provide further insight into the roles of SRF in normal development and disease. More >

Pathway: Serum Response

Induction Treatment: 20% Fetal Bovines Serum (FBS)

Glucocorticoids play an essential role in maintaining basal and stress-related homeostasis. Specifically, Glucocorticoid Receptor (GR) can act as a transcription factor by binding glucocorticoid response elements (GREs), or as a regulator of other transcription factors. GR mutations can result in glucocorticoid resistance or hypersensitivity, pathologic alterations of metabolism and excessive or suppressed immune response. More >

Pathway: Glucocorticoid receptor

Induction Treatment: Dexamethasone (DMSO),
Prednisone, Cortisone

The Estrogen receptor (ER) mediates the action of the endogenous steroid hormone, 17β-estradiol (E2). ER is synthesized as two protein forms, ERα and ERβ, which can bind to the estrogen receptor element (ERE) as either homo- or hetero- dimers. Cell proliferation and differentiation triggered by ER activation has been heavily studied in bone-wasting and breast cancer treatments. Estrogen receptors are also necessary for sexual development and reproductive function. More >

Pathway: Estrogen receptor

Induction Treatment: β-estradiol (E2)

The Androgen receptor (AR) facilitates signaling of testosterone and other androgenic hormones in the cytoplasm. Upon hormone binding, the AR translocates into the nucleus and serves as a DNA-binding activator of transcription for hormone-responsive genes. Androgen Receptor is also involved in the development of both primary and secondary male sexual characteristics and abnormal expression of AR is linked to prostate cancer. More >

Pathway: Androgen receptor

Induction Treatment: Methyltrienolone (R1881)