Header menu link for other important links
X
A novel reporter system for cyclic AMP mediated gene expression in mammalian cells based on synthetic transgene expression system
Published in Elsevier B.V.
2019
PMID: 31034821
Volume: 855
   
Pages: 56 - 64
Abstract
Cyclic AMP (cAMP)is an important second messenger that mediates various biological functions in both prokaryotes and eukaryotes. Due to the ever increasing significance in studying the function and modulation of cAMP-based signaling, it is important to develop a protein-based biosensor that reports the cAMP mediated gene expression. Based on a synthetic transgene approach, an artificial mammalian transactivator was developed by fusing a transcriptional regulatory element cAMP receptor protein (CRP)of Escherichia coli to the VP16 transactivation domain of Herpes simplex virus in a mammalian expression vector (pLA1)that activates CRP specific operator site present in a chimeric promoter (OCRP- PhCMVmin- Luciferase)in a concentration dependent manner in mammalian cells. Our results reveal that the engineered transactivator report the gene expression mediated by cAMP directly in mammalian cells and this cAMP reporter system works irrespective of Protein kinase A (PKA)- cyclic AMP response element binding protein (CREB)- cyclic AMP response element (CRE)signaling since the luciferase activity mediated by synthetic gene construct is seen even in the presence of PKA inhibitor H-89 (derived from H-8 (N-[2-(methylamino)ethyl]-5-isoquinoline-sulfonamide). Furthermore this synthetic transcription factor plays a significant role in reporting and mediating cAMP signaling in tumorigenic cells which possess an aberrant cAMP signaling due to PKA and CREB mutations. © 2019
About the journal
JournalData powered by TypesetEuropean Journal of Pharmacology
PublisherData powered by TypesetElsevier B.V.
ISSN00142999
Open AccessNo
Concepts (59)
  •  related image
    Cyclic amp
  •  related image
    Cyclic amp dependent protein kinase
  •  related image
    CYCLIC AMP RECEPTOR
  •  related image
    CYCLIC AMP RESPONSIVE ELEMENT BINDING PROTEIN
  •  related image
    Luciferase
  •  related image
    N [2 (4 BROMOCINNAMYLAMINO)ETHYL] 5 ISOQUINOLINESULFONAMIDE
  •  related image
    PROTEIN VP16
  •  related image
    CRP PROTEIN, E COLI
  •  related image
    CYCLIC AMP BINDING PROTEIN
  •  related image
    CYCLIC AMP RESPONSIVE ELEMENT BINDING PROTEIN
  •  related image
    Escherichia coli protein
  •  related image
    A-549 CELL LINE
  •  related image
    Animal cell
  •  related image
    Article
  •  related image
    Cho cell line
  •  related image
    Concentration response
  •  related image
    Controlled study
  •  related image
    CYCLIC AMP RESPONSIVE ELEMENT
  •  related image
    Escherichia coli
  •  related image
    EXPRESSION VECTOR
  •  related image
    Gene activation
  •  related image
    Gene expression system
  •  related image
    Gene mutation
  •  related image
    Hek293 cell line
  •  related image
    HERPES SIMPLEX VIRUS
  •  related image
    Human
  •  related image
    Human cell
  •  related image
    KHOS/NP CELL LINE
  •  related image
    Mammal cell
  •  related image
    Mcf-7 cell line
  •  related image
    Nonhuman
  •  related image
    Priority journal
  •  related image
    Promoter region
  •  related image
    REPORTER GENE
  •  related image
    Signal transduction
  •  related image
    Synthetic biology
  •  related image
    TRANSACTIVATION
  •  related image
    Transcription regulation
  •  related image
    TRANSGENE
  •  related image
    Animal
  •  related image
    Cell line
  •  related image
    Gene expression
  •  related image
    Genetic engineering
  •  related image
    Genetic transcription
  •  related image
    Genetics
  •  related image
    Intracellular space
  •  related image
    Metabolism
  •  related image
    Procedures
  •  related image
    REPORTER GENE
  •  related image
    TRANSGENE
  •  related image
    Animals
  •  related image
    CYCLIC AMP RECEPTOR PROTEIN
  •  related image
    CYCLIC AMP RESPONSE ELEMENT-BINDING PROTEIN
  •  related image
    Cyclic amp-dependent protein kinases
  •  related image
    Escherichia coli proteins
  •  related image
    GENES, REPORTER
  •  related image
    Humans
  •  related image
    Transcription, genetic
  •  related image
    TRANSGENES