Difference between revisions of "Part:BBa K5477024"

Latest revision as of 00:45, 2 October 2024

pRAD27-AhR - receptor module

The pSTE12-AhR receptor module is designed as a composite with AhR BBa_K3793004 is expressed under the control of the pRAD27 promoter BBa_K5477002. AhR is a ligand-activated transcription factor that binds to a variety of environmental contaminants, including polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and dioxins (1) (2). Upon binding these ligands, AhR translocates from the cytoplasm to the nucleus, where it forms a heterodimer with the Aryl hydrocarbon receptor nuclear translocator (ARNT). This complex then binds to xenobiotic response elements (XREs), initiating the transcription of detoxification genes, such as those encoding cytochrome P450 enzymes.

The pRAD27-AhR module was used for the biosensor device to detect environmental pollutants like PAHs and PCBs. Under the presence of these compounds, receptor modules including ARNT BBa_K5477025 and NCOA BBa_K5477026 will bind to XRE and initiate the transcription of NanoLuc in our reporter module BBa_K5477030. The composite part was cloned using the method of USER-cloning into YCp-L. The YCp-L plasmid is a centromeric plasmid that carries a LEU2 marker for leucine selection in yeast. Centromeric plasmids like YCp-L are low-copy vectors, typically maintained at one to two copies per cell due to the presence of a CEN sequence. This composite part was used in our device: BBa_K5477042

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 1064
Illegal SpeI site found at 289
Illegal SpeI site found at 426
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 1064
Illegal NheI site found at 869
Illegal SpeI site found at 289
Illegal SpeI site found at 426
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 1064
Illegal BglII site found at 102
Illegal BglII site found at 1879
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 1064
Illegal SpeI site found at 289
Illegal SpeI site found at 426
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 1064
Illegal SpeI site found at 289
Illegal SpeI site found at 426
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. Mandal A, Biswas N, Alam MN. Implications of xenobiotic-response element(s) and aryl hydrocarbon receptor in health and diseases. Hum Cell. 2023 Sep;36(5):1638-1655. doi: 10.1007/s13577-023-00931-5. Epub 2023 Jun 17. PMID: 37329424.

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K5477024 short</partinfo>
-The pRAD27-AhR receptor module integrates the Aryl Hydrocarbon Receptor (AhR) [https://parts.igem.org/Part:BBa_K3793004| BBa_K3793004] with the pRAD27 promoter [https://parts.igem.org/Part:BBa_K5477002| BBa_K5477002], creating a system that is responsive to both environmental toxins and cellular stress related to DNA repair processes. The pRAD27 promoter is derived from the RAD27 gene, which is involved in DNA repair and replication in yeast, particularly in the processing of Okazaki fragments during DNA synthesis.
-In this composite module, the pRAD27 promoter controls the expression of AhR, a ligand-activated transcription factor that senses and responds to environmental contaminants such as polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and dioxins (1) (2). When these toxins bind to AhR, the receptor translocates to the nucleus, where it dimerizes with the Aryl hydrocarbon receptor nuclear translocator (ARNT). This complex then binds to xenobiotic response elements (XREs) in the DNA, activating the transcription of detoxification genes, such as those coding for cytochrome P450 enzymes, which metabolize and neutralize harmful compounds.
+The pSTE12-AhR receptor module is designed as a composite with AhR [https://parts.igem.org/Part:BBa_K3793004| BBa_K3793004] is expressed under the control of the pRAD27 promoter [https://parts.igem.org/Part:BBa_K5477002| BBa_K5477002]. AhR is a ligand-activated transcription factor that binds to a variety of environmental contaminants, including polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and dioxins (1) (2). Upon binding these ligands, AhR translocates from the cytoplasm to the nucleus, where it forms a heterodimer with the Aryl hydrocarbon receptor nuclear translocator (ARNT). This complex then binds to xenobiotic response elements (XREs), initiating the transcription of detoxification genes, such as those encoding cytochrome P450 enzymes.
-The pRAD27-AhR module is used for our biosensor system for detecting environmental pollutants like PAHs and PCBs. Under the presence of these compounds, receptor modules including ARNT [https://parts.igem.org/Part:BBa_K5477025 BBa_K5477025] and NCOA [https://parts.igem.org/Part:BBa_K5477026 BBa_K5477026] will bind to XRE and initiate the transcription of NanoLuc in our reporter module [https://parts.igem.org/Part:BBa_K5477030 BBa_K5477030]. The composite part was cloned using the method of USER-cloning into YCp-L. The YCp-L plasmid is a centromeric plasmid that carries a LEU2 marker for leucine selection in yeast. Centromeric plasmids like YCp-L are low-copy vectors, typically maintained at one to two copies per cell due to the presence of a CEN sequence. This composite part was used in our device: [https://parts.igem.org/Part:BBa_K5477042 BBa_K5477042]
+The pRAD27-AhR module was used for the biosensor device to detect environmental pollutants like PAHs and PCBs. Under the presence of these compounds, receptor modules including ARNT [https://parts.igem.org/Part:BBa_K5477025 BBa_K5477025] and NCOA [https://parts.igem.org/Part:BBa_K5477026 BBa_K5477026] will bind to XRE and initiate the transcription of NanoLuc in our reporter module [https://parts.igem.org/Part:BBa_K5477030 BBa_K5477030]. The composite part was cloned using the method of USER-cloning into YCp-L. The YCp-L plasmid is a centromeric plasmid that carries a LEU2 marker for leucine selection in yeast. Centromeric plasmids like YCp-L are low-copy vectors, typically maintained at one to two copies per cell due to the presence of a CEN sequence. This composite part was used in our device: [https://parts.igem.org/Part:BBa_K5477042 BBa_K5477042]