Difference between revisions of "Part:BBa K3793005"
 
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Author: Kate Escobar, 26-09-2024, MilkClear's Wiki: https://2024.igem.wiki/ucopenhagen/
 
Author: Kate Escobar, 26-09-2024, MilkClear's Wiki: https://2024.igem.wiki/ucopenhagen/
  
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This part was used in our biosensor system to detect polycyclic aromatic hydrocarbons (PAHs), dioxin-like polychlorinated biphenyls (PCBs) and dioxin in breast milk. More information about our AhR biosensor devices, where ARNT is included, are linked here: [https://parts.igem.org/Part:BBa_K5477041 | BBa_K5477041] and [https://parts.igem.org/Part:BBa_K5477042 | BBa_K5477042].
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Aryl hydrocarbon receptor nuclear translocator (ARNT) forms heterodimerization with the Aryl hydrocarbon receptor (AhR) in the detection and response to environmental pollutants (1), particularly compounds like polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and dioxin (2) (3). Once they bind to AhR in the cytoplasm, the receptor undergoes a conformational change that exposes its nuclear localization signal, allowing it to translocate into the nucleus. There, AhR forms a complex with ARNT, and this heterodimer binds to specific DNA sequences called xenobiotic response elements (XREs) [https://parts.igem.org/Part:BBa_K5477008 BBa_K5477008] in the promoters of target genes, initiating the transcription of detoxification enzymes.
  
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This part was used to detect polycyclic aromatic hydrocarbons (PAHs), dioxin-like polychlorinated biphenyls (PCBs) and dioxin. More information about our AhR biosensor devices, where ARNT is included, are linked here: [https://parts.igem.org/Part:BBa_K5477041 | BBa_K5477041] and [https://parts.igem.org/Part:BBa_K5477042 | BBa_K5477042].
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Aryl hydrocarbon receptor nuclear translocator (ARNT) is a crucial partner of the Aryl hydrocarbon receptor (AhR) in the detection and response to environmental pollutants (1), particularly compounds like polycyclic aromatic hydrocarbons (PAHs) (2) and polychlorinated biphenyls (PCBs) (3). Once PAHs or PCBs bind to AhR in the cytoplasm, the receptor undergoes a conformational change that exposes its nuclear localization signal, allowing it to translocate into the nucleus. There, AhR forms a complex with ARNT, and this heterodimer binds to specific DNA sequences called xenobiotic response elements (XREs) [https://parts.igem.org/Part:BBa_K5477008 | BBa_K5477008] in the promoters of target genes, initiating the transcription of detoxification enzymes such as CYP1A1, which metabolizes toxic compounds into less harmful compounds.
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In our receptor module, ARNT acts as a co-regulator with AhR1 to mediate transcriptional responses to the presence of PAHs and PCBs [https://parts.igem.org/Part:BBa_K5477025 | BBa_K5477025]. Without ARNT, the AhR pathway cannot properly initiate our reporter module [https://parts.igem.org/Part:BBa_K5477030 | BBa_K5477030].
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Latest revision as of 21:13, 1 October 2024


CDS - Human AhR Nuclear Translocator (Truncated)

This sequence encodes for the truncated version of the human Aryl hydrocarbon receptor Nuclear Translocator(ARNT). ARNT is the dimerizing partner of Aryl hydrocarbon receptor (AhR). The heterodimer of AhR-ARNT function as a transcription factor in a ligand-activated fashion. Upon ligand binding to AhR, it gets translocated to the nucleus where it dimerizes with ARNT. The AhR-ARNT heterodimer binds to the binds to the DNA sequence 5'-TGCGTG-3' within the dioxin response element (DRE) or xenobiotic response element (XRE) of target gene promoters and activates their transcription.

The part contains the coding sequence for a truncated version of ARNT containing the the basic helix-loop-helix, PAS-A and PAS-B domains. The CDS has been codon optimized to remove any type IIS restriction enzyme sites. An N-Terminal 6x HIS tag is added to the CDS.


MilkClear iGEM UCopenhagen 2024

Author: Kate Escobar, 26-09-2024, MilkClear's Wiki: https://2024.igem.wiki/ucopenhagen/

Aryl hydrocarbon receptor nuclear translocator (ARNT) forms heterodimerization with the Aryl hydrocarbon receptor (AhR) in the detection and response to environmental pollutants (1), particularly compounds like polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and dioxin (2) (3). Once they bind to AhR in the cytoplasm, the receptor undergoes a conformational change that exposes its nuclear localization signal, allowing it to translocate into the nucleus. There, AhR forms a complex with ARNT, and this heterodimer binds to specific DNA sequences called xenobiotic response elements (XREs) BBa_K5477008 in the promoters of target genes, initiating the transcription of detoxification enzymes.

This part was used to detect polycyclic aromatic hydrocarbons (PAHs), dioxin-like polychlorinated biphenyls (PCBs) and dioxin. More information about our AhR biosensor devices, where ARNT is included, are linked here: | BBa_K5477041 and | BBa_K5477042.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 104
    Illegal EcoRI site found at 1210
    Illegal SpeI site found at 1054
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 104
    Illegal EcoRI site found at 1210
    Illegal SpeI site found at 1054
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 104
    Illegal EcoRI site found at 1210
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 104
    Illegal EcoRI site found at 1210
    Illegal SpeI site found at 1054
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 104
    Illegal EcoRI site found at 1210
    Illegal SpeI site found at 1054
  • 1000
    COMPATIBLE WITH RFC[1000]


References

1. Carambia, A., Schuran, F.A. The aryl hydrocarbon receptor in liver inflammation. Semin Immunopathol 43, 563–575 (2021). https://doi.org/10.1007/s00281-021-00867-8

2. Goedtke L, Sprenger H, Hofmann U, Schmidt FF, Hammer HS, Zanger UM, Poetz O, Seidel A, Braeuning A, Hessel-Pras S. Polycyclic Aromatic Hydrocarbons Activate the Aryl Hydrocarbon Receptor and the Constitutive Androstane Receptor to Regulate Xenobiotic Metabolism in Human Liver Cells. Int J Mol Sci. 2020 Dec 31;22(1):372. doi: 10.3390/ijms22010372. PMID: 33396476; PMCID: PMC7796163.

3. Kafafi SA, Afeefy HY, Ali AH, Said HK, Kafafi AG. Binding of polychlorinated biphenyls to the aryl hydrocarbon receptor. Environ Health Perspect. 1993 Oct;101(5):422-8. doi: 10.1289/ehp.93101422. PMID: 8119253; PMCID: PMC1519849.