Part:BBa_K4674006
Tetrahedral ssDNA with cis-auto splicing motif
Over the past few years, the swift advancement of DNA nanotechnology has brought forth fresh perspectives for the detection and therapy of cancer. These nanostructures, assembled from single-stranded DNA, exhibit several features that make them suitable candidates for innovative cancer detection strategies.
One of the main advantages of DNA tetrahedron-based approaches in cancer detection is their high programmability and specificity. These structures can be precisely designed to carry various functional components, including ligands. This allows us to create tailored probes capable of targeting specific cancer-related biomarkers with a high degree of accuracy. Additionally, the DNA tetrahedron's inherent stability and rigidity contribute to the stability of the detection platform. This stability is essential for maintaining the integrity of the assembled nanostructure during various stages of the detection process, including sample preparation, target binding, and signal readout.
In summary, DNA tetrahedron structures offer a promising platform for the development of innovative cancer detection methods. Their programmability, versatility, and ability to carry multiple functional components make them well-suited for designing highly sensitive and specific assays that hold the potential to contribute significantly to improved cancer detection and patient outcomes.
The design of DNA tetrahedron assembled from trtrahedral ssDNA
The folding pathway of the tetrahedral ssDNA is illustrated in the figure below. Among six edges, five are composed of 21 bp double helix. The last edge is a “twin double helices” to compensate the necessary of reverse polarity of complementary DNA strands. The 5’ terminus of tetrahedral ssDNA starts at one endpoint, and the PBSII and Zif268 binding motifs are located at the farest two edges, respectively (Conrado, R. J., et al. 2011). The PBSII and Zif268 binding motifs are applied to anchor tetrahedrons on the wall of main chamber in CTC-FAST device though interaction with PBSII and Zif268 proteins.
Since the commercial production of tetrahedral ssDNA is hampered by its highly complementary sequence, we decided to apply the RCR mechanism to gererate the tetrahedral ssDNA (see below section for details). The products from RCR will be circular ssDNA, therefore, we decided to fuse the cis-auto splicing sequence at the 5’ terminus tetrahedral ssDNA. After auto-splicing, the RCR generated circular ssDNA will fold into tetradron, and a 18 nucleotides overhang, which are the residues after splicing, will left at 3’ terminus. The 3’ terminus of FA-conjugated adaptor ssDNA will be complement to this overhang, and the 5’ terminus will contain the trans-auto splicing sequence. Finally, the exogenous ssDNA complementary to the trans-auto slicng sequence could be added to cleave the adaptor ssDNA and release the captured CTCs.
Fig. The design design of DNA tetrahedron assembled from tetrahhedral ssDNA.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 107
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 107
Illegal NheI site found at 185 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 107
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 107
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 107
Illegal NgoMIV site found at 166 - 1000COMPATIBLE WITH RFC[1000]
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