Team:REC-CHENNAI/Parts-Construction
PART CONSTRUCTION
This part is a 47-base containing short hairpin RNA (shRNA) synthesized in-vitro, with 4 bases in the tight hairpin loop and other 43 bases in the stem. The silencing sequence 5’-GCAUUGGGCGCUGAUUCUU-3’ is present in the stem region, which silences survivin mRNA through the mechanism of interference.
This part is a 25-base containing small interfering RNA (siRNA) synthesized in-vitro, with 3-bp mediated self-complimentarity formed between their two termini, introduced to form an shRNA-like structure. This forms a structure of short stem containing 3-bp, and a large loop containing 22-bp, which houses the silencing sequence. The 5’ end of the stem region is conjugated with the fluorophore 6-FAM (which has an excitation wavelength of 495 nm and an emission wavelength of 520nm) and the 3’ end is conjugated with the quencher IB-FQ.
This part is the coding sequence of the chimeric CPP formed by BP100 and the polycationic peptide (KH)9. It is proven to deliver proteins and nucleic acids into the plant and prokaryotic cells. The polycationic peptide is an efficient nucleic acid carrier and has the ability to promote buffering effect on the pre-lysosomal vesicle. The cargo being negatively charged interacts with the polycationic end of the peptide through ionic interactions, whereas the BP100 region mediates cellular entry by interfering with the actin dynamics of the cell.
