Difference between revisions of "Part:BBa K5477025"

Latest revision as of 00:52, 2 October 2024

pRET2 - ARNT receptor module

The pRET2-ARNT receptor module is a composite part designed to control the expression of ARNT (Aryl hydrocarbon receptor nuclear translocator) | BBa_K3793005 under the regulation of the pRET2 promoter BBa_K5477000. ARNT forms heterodimerization with the Aryl hydrocarbon receptor (AhR) in the nucleus. This heterodimer regulate gene transcription in response to xenobiotics (1) (2). This complex then binds to xenobiotic response elements (XREs) in the promoter regions of detoxification genes (1) (2) (3) (4).

This module supports the AhR-mediated response to toxic ligands like PAHs, dioxins and PCBs by binding to the XRE in the reporter module facilitating the transcription of NanoLuc BBa_K5477030. The composite part was cloned using the method of USER-cloning into YCp-H. YCp-H is a centromeric plasmid used in yeast that includes a HIS3 marker, allowing for selection in histidine auxotrophic yeast strains. Like other CEN plasmids, YCp-H contains a CEN sequence, ensuring that the plasmid replicates and segregates similarly to yeast chromosomes. This results in a low copy number (typically one to two copies per cell), providing stable maintenance of the plasmid. This composite part was used in the following devices: BBa_K5477041 and BBa_K5477042

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 810
Illegal EcoRI site found at 1916
Illegal SpeI site found at 1760
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 810
Illegal EcoRI site found at 1916
Illegal NheI site found at 182
Illegal SpeI site found at 1760
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 810
Illegal EcoRI site found at 1916
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 810
Illegal EcoRI site found at 1916
Illegal SpeI site found at 1760
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 810
Illegal EcoRI site found at 1916
Illegal SpeI site found at 1760
Illegal AgeI site found at 37
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.

4. Huang G, Elferink CJ. A novel nonconsensus xenobiotic response element capable of mediating aryl hydrocarbon receptor-dependent gene expression. Mol Pharmacol. 2012 Mar;81(3):338–47.

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K5477025 short</partinfo>
-The pRET2-ARNT receptor module is a composite part designed to control the expression of ARNT (Aryl hydrocarbon receptor nuclear translocator) [https://parts.igem.org/Part:BBa_K3793005 | BBa_K3793005] under the regulation of the pRET2 promoter [https://parts.igem.org/Part:BBa_K5477000| BBa_K5477000]. ARNT is a crucial partner in the Aryl hydrocarbon receptor (AhR) pathway, functioning as a heterodimerization partner that allows AhR to regulate gene transcription in response to environmental toxins (1) (2). When AhR binds to toxic compounds like polycyclic aromatic hydrocarbons (PAHs) or polychlorinated biphenyls (PCBs), it translocates into the nucleus and requires ARNT to form a functional transcriptional complex. This complex then binds to xenobiotic response elements (XREs) in the promoter regions of detoxification genes, such as those encoding cytochrome P450 enzymes, initiating the breakdown and clearance of harmful substances (1) (2) (3).
+The pRET2-ARNT receptor module is a composite part designed to control the expression of ARNT (Aryl hydrocarbon receptor nuclear translocator) [https://parts.igem.org/Part:BBa_K3793005 | BBa_K3793005] under the regulation of the pRET2 promoter [https://parts.igem.org/Part:BBa_K5477000| BBa_K5477000]. ARNT forms heterodimerization with the Aryl hydrocarbon receptor (AhR) in the nucleus. This heterodimer regulate gene transcription in response to xenobiotics (1) (2). This complex then binds to xenobiotic response elements (XREs) in the promoter regions of detoxification genes (1) (2) (3) (4).
-This module supports the AhR-mediated response to toxic ligands like PAHs, dioxins and PCBs by binding to the XRE in our reporter module facilitating the transcription of NanoLuc [https://parts.igem.org/Part:BBa_K5477030 BBa_K5477030]. The composite part was cloned using the method of USER-cloning into YCp-H. YCp-H is a centromeric plasmid used in yeast that includes a HIS3 marker, allowing for selection in histidine auxotrophic yeast strains. Like other CEN plasmids, YCp-H contains a CEN sequence, ensuring that the plasmid replicates and segregates similarly to yeast chromosomes. This results in a low copy number (typically one to two copies per cell), providing stable maintenance of the plasmid. This composite part was used in our devices: [https://parts.igem.org/Part:BBa_K5477041 BBa_K5477041] and [https://parts.igem.org/Part:BBa_K5477042 BBa_K5477042]
+This module supports the AhR-mediated response to toxic ligands like PAHs, dioxins and PCBs by binding to the XRE in the reporter module facilitating the transcription of NanoLuc [https://parts.igem.org/Part:BBa_K5477030 BBa_K5477030]. The composite part was cloned using the method of USER-cloning into YCp-H. YCp-H is a centromeric plasmid used in yeast that includes a HIS3 marker, allowing for selection in histidine auxotrophic yeast strains. Like other CEN plasmids, YCp-H contains a CEN sequence, ensuring that the plasmid replicates and segregates similarly to yeast chromosomes. This results in a low copy number (typically one to two copies per cell), providing stable maintenance of the plasmid. This composite part was used in the following devices: [https://parts.igem.org/Part:BBa_K5477041 BBa_K5477041] and [https://parts.igem.org/Part:BBa_K5477042 BBa_K5477042]
@@ Line 28: / Line 28: @@
 . 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.
+. Huang G, Elferink CJ. A novel nonconsensus xenobiotic response element capable of mediating aryl hydrocarbon receptor-dependent gene expression. Mol Pharmacol. 2012 Mar;81(3):338–47.