Parts
Parts Library
Our Parts Library provides a complete collection of all biological parts used in the LEGO system, including both basic parts and composite parts. Based on their functional roles, these parts are categorized into three groups: optogenetic system validation elements, light-responsive activator of the LEGO system, and dual-hormone expression vector of the LEGO system.
Optogenetic system validation elements
The optogenetic system validation elements include components used in experiments validating the sustained nuclear localization of the iLID-sspB system, blue-light responsiveness, and bioluminescence-induced activation. These parts employ localization sequences to target iLID and sspB to specific subcellular compartments (e.g., nucleus, cytoplasm, plasma membrane), while fused fluorescent proteins enable direct visualization of protein localization and dynamics under microscopy. Selected components also incorporate NanoLuc luciferase for generating endogenous bioluminescence. The efficient assembly of these parts provides a concise and reliable optogenetic toolkit for validating the applicability of light-responsive systems.
| Part Number | Part Name | Type | Composition | Status | Registry |
|---|---|---|---|---|---|
| BBa_25SVZLG1 | iLID improved light-induced dimer | Coding | Basic Part | Uploaded by WLSA Shanghai Academy but not novel | Link |
| BBa_25CKWQSO | SspB nano | Coding | Basic Part | Uploaded by WLSA Shanghai Academy but not novel | Link |
| BBa_K5036002 | NLS | Signaling | Basic Part | Existing | Link |
| BBa_25B1YMN7 | NLyn membrane targeting sequence | Coding | Basic Part | New | Link |
| BBa_25UVDTT5 | miRFP670nano3 deep-red fluorescent protein | Coding | Basic Part | Uploaded by WLSA Shanghai Academy but not novel | Link |
| BBa_25XIDIXX | mCherry | Coding | Basic Part | Existing | Link |
| BBa_25AWEP97 | Nluc | Coding | Basic Part | New | Link |
| BBa_25T0DJLM | NLS-miRFP670nano3-SspB | Coding | Composite Part | New | Link |
| BBa_257KZSX5 | NLyn-miRFP670nano3-SspB | Coding | Composite Part | New | Link |
| BBa_25ZVZJXP | NLuc-iLID-mCherry | Coding | Composite Part | New | Link |
| BBa_25FPKXWO | NLS-Nluc-iLID-mCherry | Coding | Composite Part | New | Link |
Light-responsive activator of the LEGO system
The components in this section were assembled to create the optogenetic transcriptional activator of the LEGO system, incorporating functional modules including nuclear localization, light-responsive dimerization, DNA-binding, and transcriptional activation domains. These modules were fused with fluorescent proteins to enable spatial visualization. The final version of the optogenetic activator also integrated NanoLuc luciferase, enabling potential activation via endogenous bioluminescence. All components were consolidated into a pair of light-inducible dimerizing elements and co-expressed from a single vector through an IRES-mediated bicistronic design, resulting in a minimal-function optogenetic transcriptional activator.
| Part Number | Part Name | Type | Composition | Status | Registry |
|---|---|---|---|---|---|
| BBa_25EWENZZ | Gal4DBD Gal4 DNA binding domain | Protein_Domain | Basic Part | New | Link |
| BBa_J176013 | VP64 | Protein_Domain | Basic Part | Existing | Link |
| BBa_K3338004 | Internal ribosome entry site (IRES) for use in eukaryotic cells | RBS | Basic Part | Existing | Link |
| BBa_25XW3FSE | NLS-Nluc-iLID-VP64-IRES-NLS-Gal4DBD-miRFP670nano3-SspB light inducible transcription activator | Coding | Composite Part | New | Link |
Dual-hormone expression vector of the LEGO system
The core components of this section consist of our engineered human insulin and bioactive GLP-1 constructs, designed for expression and secretion in mammalian cells. The dual-hormone genes and a fluorescent reporter gene mCherry are linked via IRES, all driven by a 5×UAS-guided Hsp70 promoter that provides a specific binding scaffold for the optogenetic activator.
| Part Number | Part Name | Type | Composition | Status | Registry |
|---|---|---|---|---|---|
| BBa_254SX1VP | Furin Sites-Inserted Human Preproinsulin | Coding | Basic Part | New | Link |
| BBa_259V4OIU | Human GLP-1 (7-37) (A8G) Fused with Extendin-4 Secretion Signal | Coding | Basic Part | New | Link |
| BBa_25KHXHFS | Hsp70 promoter | Prometer | Basic Part | New | Link |
| BBa_K5182008 | The upstream activating sequence (UAS) is a region that interacts with the transcriptional regulator | Coding | Basic Part | Existing | Link |
| BBa_251FTTH1 | UAS-Hsp70 inducible promoter | Prometer | Composite Part | New | Link |
| BBa_25EIL13E | Ins-IRES-GLP1-IRES-mCherry for human preproinsulin and human GLP1 (7-37) (A8G) expression | Coding | Composite Part | New | Link |