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Overview

We submitted 19 well-documented BioBrick parts, including all those used in our project. This collection comprises 9 basic parts (see Basic Parts page) and 10 composite parts (see Composite parts page). We provided detailed characterization data for several important parts. All parts used in our project had been submitted to the iGEM Registry.
Our favorite basic part is MTNR1A (BBa_K5267001), and our favorite composite part is Pmin_4*CRE->IgK->Nluc->bGH_polyA (BBa_K5267040).

Figure 1. Building a mammalian cell-based drug screening platform based on MTNR1A

Favorite basic Part (BBa_K5267001)

MTNR1A( BBa_K5267001), is one of the most crucial part in our parts collection, as it encodes the melatonin receptor MTNR1A and act as a sensor to respond to compounds capable of effectively activating melatonin receptor (agonist). It lays the foundational basis for cell-based screening platform and subsequent experiments in our project aimed at identifying potential melatonin receptor agonist.

Figure 2 Concrete schematic diagram of BBa_K5267040 composite part

Favorite Composite Part(BBa_K5267040)

Pmin_4*CRE->IgK->Nluc->bGH_polyA(BBa_K5267040) functions as a sensor for cAMP/PKA/CREB pathway in mammalian cell. Upon pathway activation, mammalian cells transfected with this composite part will respond to the activation signals by secreting NanoLuc luciferase into the cell medium.
By co-transfecting HEK-293T cell lines with one plasmid carrying the construct PCMV->MTNR1A->bGH_polyA (Part: BBa_K5267047) and another carrying the construct P_4*CRE->IgK->Nluc->bGH_polyA (Part: BBa_K5267040), we can build a cell-based high-throughput drug screening platform. This platform allows for the simultaneous assessment of multiple cAMP/PKA/CREB pathway-activating drug candidates through resource-efficient and straightforward experiments.

For more detailed information about our parts, we highly recommend visiting the specific part list.

Parts List

BBa_K5267001	MTNR1A	Encoding melatonin receptor MTNR1a (MTNR1A), a G protein-coupled receptor (GPCR) primarily coupled to the Gi/o protein family and activates the downstream signaling when receives the stimulation of melatonin or its analogues.	Coding	1050bp
BBa_K5267002	MTNR1B	Encoding melatonin receptor MTNR1b (MTNR1B), a G protein-coupled receptor (GPCR) primarily coupled to the Gi/o protein family and activates the downstream signaling when receives the stimulation of melatonin or its analogues.	Coding	1089bp
BBa_K5267004	Pmin_4*CRE promoter	A regulated promoter made up of 4 copies of CRE (cAMP Response Element) and a minimal promoter that response to the activation of CREB (cAMP Response Element Binding Protein). Activated CREB binds to the CREs and activate the transcription of the downstream genes.	Regulatory	240bp
BBa_K5267005	Pmin_5*CRE promoter	A regulated promoter made up of 5 copies of CRE (cAMP Response Element) and a minimal promoter that response to the activation of CREB (cAMP Response Element Binding Protein). Activated CREB binds to the CREs and activate the transcription of the downstream genes.	Regulatory	255bp
BBa_K5267006	Pmin_6*CRE promoter	A regulated promoter made up of 6 copies of CRE (cAMP Response Element) and a minimal promoter that response to the activation of CREB (cAMP Response Element Binding Protein). Activated CREB binds to the CREs and activate the transcription of the downstream genes.	Regulatory	270bp
BBa_K5267007	Pmin_1*NFAT promoter	A regulated promoter made up of 1 copies of NFAT binding sites and a minimal promoter that response to the dephosphorylation of NFAT protein. Dephosphorylated NFAT proteins undergo nuclear translocation and bind to NFAT binding sites, activating the transcription of the downstream genes.	Regulatory	69bp
BBa_K5267008	Pmin_5*NFAT promoter	A regulated promoter made up of 5 copies of NFAT binding sites and a minimal promoter that response to the dephosphorylation of NFAT protein. Dephosphorylated NFAT proteins undergo nuclear translocation and bind to NFAT binding sites, activating the transcription of the downstream genes.	Regulatory	191bp
BBa_K5267009	Pmin_6*NFAT promoter	A regulated promoter made up of 6 copies of NFAT binding sites and a minimal promoter that response to the dephosphorylation of NFAT protein. Dephosphorylated NFAT proteins undergo nuclear translocation and bind to NFAT binding sites, activating the transcription of the downstream genes.	Regulatory	219bp
BBa_K5267010	Pmin_7*NFAT promoter	A regulated promoter made up of 7 copies of NFAT binding sites and a minimal promoter that response to the dephosphorylation of NFAT protein. Dephosphorylated NFAT proteins undergo nuclear translocation and bind to NFAT binding sites, activating the transcription of the downstream genes.	Regulatory	249bp
BBa_K5267040	Pmin_4*CRE->IgK->Nluc->bGH_polyA	Using Nanoluc to characterize the MTNR1A-mediated activation of cAMP-PKA signaling pathway	Composite	944bp
BBa_K5267041	Pmin_5*CRE->IgK->Nluc->bGH_polyA	Using Nanoluc to characterize the MTNR1A-mediated activation of cAMP-PKA signaling pathway	Composite	1082bp
BBa_K5267042	Pmin_6*CRE->IgK->Nluc->bGH_polyA	Using Nanoluc to characterize the MTNR1A-mediated activation of cAMP-PKA signaling pathway	Composite	1097bp
BBa_K5267043	Pmin_1*NFAT->IgK->Nluc->bGH_polyA	Using Nanoluc to characterize the MTNR1A-mediated activation of Ca²⁺ signaling pathway	Composite	896bp
BBa_K5267044	Pmin_5*NFAT->IgK->Nluc->bGH_polyA	Using Nanoluc to characterize the MTNR1A-mediated activation of Ca²⁺ signaling pathway	Composite	1018bp
BBa_K5267045	Pmin_6*NFAT->IgK->Nluc->bGH_polyA	Using Nanoluc to characterize the MTNR1A-mediated activation of Ca²⁺ signaling pathway	Composite	1046bp
BBa_K5267046	Pmin_7*NFAT->IgK->Nluc->bGH_polyA	Using Nanoluc to characterize the MTNR1A-mediated activation of Ca²⁺ signaling pathway	Composite	1076bp
BBa_K5267047	P_PCMV->MTNR1A->bGH_polyA	Using strong promoter to initiate MTNR1A expression	Composite	19099bp
BBa_K5267048	P_PCMV->MTNR1B->bGH_polyA	Using strong promoter to initiate MTNR1B expression	Composite	1949bp
BBa_K5267049	P_min->IgK->Nluc->bGH_polyA	A minimal TATA-box promoter (Pmin) initiate the expression of secreted Nanoluc	Composite	859bp