Optogenetics was achieved for the biosynthetic process's control. Optogenetics extended the “Blue light-dependent interaction."[1] Compared to traditional chemotherapy， photoswitch has better controllability. Photoswitch become an essential tool in synthetic biology and targeted treatment of diseases， for example, cancer， diabetes, neurological disorders, etc. Among cancers using CAR-T treatment， blue light-dependent interaction Can be effectively optimized see Fig. 1. We improved the original blue light-dependent interaction “CRY2/CIB1”[2][3] and Improved “CRY2(GFP)/SPA1”[4]. The experiment compared the new and the old blue light-dependent interaction's sensitivity and strength.
Fig.1 CAR-T cells enable safe and controlled cellular immunotherapy
l CAR-T
Car-T treats acute and chronic lymphoblastic leukemia and other diseases in patients with malignant tumors. It is clinically targeted at tumors—a new type of immune cell precision targeting therapy technology. However, significant side effects can result in headaches, confusion, and delirium, among other neural changes.
l Cryptochromes
Cryptochromes are blue light receptors that mediate circadian rhythm and magnetic sensing in various organisms.[5] A typical cryptochrome consists of a conserved photolyase homology region domain and a varying carboxyl-terminal extension across species.[6]
l Research status of blue light-dependent interaction
Specifically, we discover that the blue-light-induced opening of carboxyl-terminal extension in C. reinhardtii animal-like cryptochrome can structurally facilitate its interaction with the Rhythm Of protein. Our finding is made possible by two technical advances. Using the single-molecule resonance energy transfer technique, we directly observe the displacement of carboxyl-terminal extension by about 15 Å upon blue light excitation. Combining structure prediction and solution X-ray scattering methods, we propose plausible structures of full-length cryptochrome under dark and light conditions. The systems provide the molecular basis for light active conformational changes of cryptochrome and downstream regulatory functions.[7]
l Advantage
² The precision of targeted therapy is improved, the accuracy of targeted therapy is enhanced, and the fault tolerance rate of the treatment is reduced.
² The damage to cells is significantly reduced.
² It does not change the composition of the medium, easy to automate production
Fug.2 Process of blue light-dependent interaction
The PCR turns on the gene segment, then restriction enzyme digestion (this helps process in the next step). Homologous recombination lets gene segment structure with plasmid. The process of heat shock conversion makes plasmid into E. coli. We are rocking inoculation to plasmid isolation. Yeast mating transfers plasmid into yeast, becomes yeast culture, and assays for β-galactosidase activity.
Future application and goal
² Combining blue light switch and CAR-T, precision medicine
² Optimize CAR-T
² It is used in severe diseases
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[2] Emery, P., So, W. V., Kaneko, M., Hall, J. C. & Rosbash, M. CRY, a Drosophila clock and light-regulated cryptochrome, major contributor to circadian rhythm resetting and photosensitivity. Cell 95, 669–679 (1998).
[3] Stanewsky, R. et al. The cryb mutation identifies cryptochrome as a circadian photoreceptor in Drosophila—cell 95, 681–692 (1998).
[4] Liu, B., Zuo, Z., Liu, H., Liu, X. & Lin, C. Arabidopsis cryptochrome one interacts with SPA1 to suppress COP1 activity in response to blue light—genes Dev. 25, 1029–1034 (2011).
[5] Chaves, Inês et al. “The cryptochromes: blue light photoreceptors in plants and animals.” Annual review of plant biology vol. 62 (2011): 335-64. doi:10.1146/annurev-arplant-042110-103759
[6] Dodson, C. A., Hore, P. J. & Wallace, M. I. A radical sense of direction: signaling and mechanism in cryptochrome magnetoreception. Trends Biochem. Sci. 38, 435–446 (2013).
[7] Communications Biology volume 5, Article number: 1103 (2022)