Part:BBa_K5073039:Design
CD28 TM: Transmembrane region of T-cell-surface glycoprotein CD28
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
During the detailed design of CD28TM, several key factors were considered to ensure its functionality and efficacy. Firstly, to ensure that the selected CD28 gene sequence is complete and can be efficiently expressed in the target host cells, the nucleotide sequence was optimized to improve translation efficiency, including a suitable RBS and promoter. In addition, select suitable plasmid vectors with good replication ability and stability, and include effective selection markers to screen for successfully transfected cells. The design also needs to ensure that the transmembrane structural domains of CD28TM are intact to ensure that it can positively localise on the cell membrane and effectively participate in cellular signal transduction and interactions. At the same time, the immunogenicity of CD28TM is assessed and the sequence is moderately mutated or optimized to reduce the immune response. In subsequent functional validation experiments, ensure that CD28TM works efficiently and synergistically with other components of the CAR, such as scFv and terminators. Finally, synthetic biology specifications were followed to ensure that the designed sequence met safety and operability requirements. These design considerations enable CD28TM to be efficiently expressed in CAR-T cells and to perform important co-stimulatory functions, thereby improving therapeutic efficacy.
Source
CD28TM is derived from the CD28 gene in the human genome. The CD28 gene is located on human chromosome 2 and encodes an important co-stimulatory molecule that is widely expressed in T-cells and other immune cells. The CD28 protein plays a key role in promoting T-cell activation and proliferation by binding to the ligands, CD80 and CD86, on the surface of antigen-presenting cells (APCs). This gene sequence can be modified by gene cloning and synthetic biology techniques to suit different experimental needs and application scenarios. In synthetic biology projects, researchers usually use relevant plasmid vectors to integrate CD28TM into chimeric antigen receptor (CAR) systems to enhance the anti-tumor effect and functional persistence of T cells.