Team:Hong Kong HKU/Part Collection
Part Collection
Best Part Collection
I. Nano Drug Carrier (NDC) w/ multiple aptamers (NDC-MA) for dual tumour-specific drug delivery system in Salmonella Typhimurium
BBa_K3063041 (Best New Basic Part)
This BioBrick encodes Strand 4-SYL3C. The aptamer, attached to this strand, SYL3C, could target the EpCAM (Epithelial cell adhesion molecule) on cancer stem cells. With the aptamer encoded by this part, our DNA nanostructure could recognise and bind to the EpCAM on cancer stem cell and thus increase the delivery efficiency of our nanostructure to cancer stem cell and reduce the stemness by unloading the Doxorubicin intercalating in the nanostructure.
BBa_K3063012
This BioBrick encodes Strand 1-ST1. The aptamer attached to this strand, ST1, could target and bind to the cell surface antigens of Salmonella Typhi for NDC attachment.
BBa_K3063022
This BioBrick encodes Strand 2-APT33. The aptamer attached to this strand, APT33, could also target and bind to the cell surface antigens of Salmonella Typhi for NDC attachment.
BBa_K3063031
This BioBrick encodes Strand 3-AS. The aptamer, attached to this strand, AS1411, could target the nucleolin on cancer cell surface for higher specificity.
BBa_K30630X0*
This BioBrick encodes Strand X-FA* which are strands with free aptamer. A special site is designed and inserted to this BioBrick for addition for desired aptamer using golden gate assembly. It is also a part of the tetrahedron body with aptamers of interest.
BBa_K30630X1*
This BioBrick encodes Strand X-AS*, a modified version of strand 1-FA. The aptamer, attached to this strand, AS1411, could target the nucleolin on cancer cell surface for higher specificity.
The interchangeable aptamer system
*where X represents the number of corresponding strand. E.g. BBa_K3063010 encodes Strand 1-FA
Interchangeable Aptamer System
To verify the interchangeable aptamer system, a mixture of short double stranded DNA oligo with different inserted restriction site and sticky ends are put into reaction with our construct. To verify the specific insertion of only a single type of fragment with specific sticky end, a restriction digestion is carried out after golden gate reaction.
Experiment shows the different sticky ends designed are incompatible and a single reaction could be carrier out for easy aptamer insertion into four different biobricks in a single reaction, allow the production of totally different aptamer presentation on DNA nanostructure in a single reaction. All replicates (5 groups, n = 4) shows correct insertion of specific fragments in fragment mixture.
The system can produce DNA nanostructures with different aptamers, that can bind to different cancer targets, allowing DNA nanostructure to bind to different bacterial carriers. The whole collection allows easy production of DNA nanostructure in vivo with different properties.
For proof of the functionality of the composite parts and basic parts mentioned, please refer to In Vivo Nanostructure Synthesis
Other Part Collections
II. Flagella Production Boost
BBa_K3063000: flhDC transcription factors (Salmonella Typhimurium)
This BioBrick encodes the flhD and flhC (flhDC) transcription factors. Upon expression of the transcription factors, it would up-regulate the expression of flagella in Salmonella Typhimurium.
BBa_K3063111: flhDC transcriptions factors (Salmonella Typhimurium) with lac promoter
This BioBrick encodes flhDC transcription factor with a constitutive lac promoter for use in Salmonella Typhimurium. It leads to expression of flhD and flhC transcription factors which up-regulates flagella production in S. Typhimurium and facilitates locomotion.
Other Part Collections
III. Artificial miRNA silencing cassette
BBa_K3063934: GFP-eSIBR cassette with STAT3 SOX4 SOX9 amiRNA construct (Best New Composite Part)
This BioBrick encodes an array of constructs. First, it encodes three artificial pre-miRNA stem loops targeting STAT3, SOX4 and SOX9. Secondly, it contained two promoter, CMV and T7, which respectively could drive transcription in mammalian cells and bacterial cells. Thirdly, it contains a f1 origin which allows the plasmid be replicating in mammalian cells.
BBa_K3063900: eSIBR cassette with amiRNA insertion site (compatible with original eSIBR plasmid)
This BioBrick allows the insertion of amiRNA into eSIBR (EGFP coding region followed by enhanced synthetic inhibitory BIC/miR-155 RNA) cassette. It allows the insertion of amiRNA sequence using golden gate cloning or standard cloning method with BpiI site, which is a type II restriction enzyme. The part is designed with an addition of SalI restriction site to the 5', and XhoI to the 3' end of the eSIBR cassette. So not only could the eSIBR cassette be assembled using the biobrick assembly methods, it could also incorporate the fragments into GFP-eSIBR expression vector stepwise using different assembly method.
BBa_K3063901: STAT3
This BioBrick encodes amiRNA that silences signal transducer and activator of transcription 3 (STAT3), a member of the STAT protein family for maintaining cancer stemness. STAT3-mediated signaling is implicated in regulating the undifferentiated phenotype of stem cells and suppressing antitumor immunity, while in vitro silencing is capable of inducing G2/M arrest in a variety of cancer cell types.
BBa_K3063902: SOX4
This BioBrick encodes amiRNA that silences SRY (sex determining region Y)-box 4 (SOX4), a gene conferring cancer stem cell (CSC) properties. SOX4, whose expression initiated by STAT3, orchestrates epithelIal-to-mesenchymal transition (EMT) through upregulating a series of EMT-inducing transcription factors, which then enhances cellular growth as well as transformation. SOX4 knockdown can effectively reduce the activation of transforming growth factor-beta (TGF-β) signaling, attenuating the invasion of cancer cells.
BBa_K3063903: SOX9
This BioBrick encodes amiRNA that silences SRY (sex determining region Y)-box 9 (SOX9), a gene promoting cancer metastasis and driving cancer cell growth. Sox9 has long been known to overexpress in tumor, in particular, cancer stem cells. It maintains self-renewal capacity of CSCs through constitutive activation of Notch signalling pathway. Metastatic cells show further elevation of Sox9 than non-metastatic ones, indicating the role of the gene in cancer progression, possibly via Wnt/β-catenin signaling.
BBa_K3063904: STAT3 & SOX4
This BioBrick encodes both STAT3- and SOX4-targeting amiRNAs that silence their corresponding gene expression.
BBa_K3063905: STAT3, SOX4, SOX9
The BioBrick encodes both STAT3-, SOX4- and SOX9-targeting amiRNA that silence their corresponding gene expression.
BBa_K3063987: eSIBR casette with amiRNA insertion sites
This BioBrick allows the insertion of amiRNA into eSIBR (EGFP coding region followed by enhanced synthetic inhibitory BIC/miR-155 RNA) cassette. It allows the insertion of amiRNA sequence using golden gate cloning or standard cloning method with BpiI site, which is a type II restriction enzyme.