<link>https://pages.htgaa.org/2026a/beyza-cennet-batir/projects/individual-final-project/beyza_dehydrin_cryoprotection/index.html</link><description>Dehydrin-Inspired Synthetic Proteins as Cryoprotective Agents for Cell Preservation: A Synthetic Biology Approach Author: Beyza Course: How to Grow (Almost) Anything — HTGAA File: ./projects/beyza_dehydrin_cryoprotection.md Abstract Cryopreservation is a cornerstone of biomedical research, cell therapy manufacturing, and biobanking, yet current cryoprotective agents (CPAs) such as dimethyl sulfoxide (DMSO) carry significant cytotoxicity risks and are poorly tolerated by sensitive cell types including stem cells and primary neurons. Nature has evolved elegant molecular solutions to freezing stress: dehydrins (DHNs), a class of intrinsically disordered proteins (IDPs) found in desiccation-tolerant plants and certain extremophilic organisms, accumulate during cold and drought stress and protect cellular membranes and proteins from freeze-induced damage. Despite decades of characterization in plant biology, dehydrin-inspired synthetic proteins have not been systematically engineered and tested as exogenous CPAs for mammalian cell preservation. This project proposes to design, synthesize, and functionally characterize three synthetic dehydrin variants — DHN-K1 (single K-segment), DHN-K2S (two K-segments plus one S-segment), and DHN-K2S-ΔS (two K-segments, S-segment deleted) — to dissect the structural determinants of cryoprotective activity. All three constructs will be codon-optimized for E. coli BL21(DE3) expression, synthesized as whole plasmids by Twist Bioscience, and expressed with N-terminal His₆-tags for affinity purification. Cryoprotective efficacy will be measured using an automated MTT cell viability assay in 384-well format on the Spark Plate Reader at Ginkgo Bioworks. The central hypothesis is that K-segment copy number positively correlates with cryoprotective activity, while the S-segment modulates but is not essential for function. This work establishes a scalable, automation-compatible pipeline for engineering next-generation protein-based CPAs with direct applications in regenerative medicine, biobanking, and cell therapy manufacturing.</description><generator>Hugo</generator><language>en</language><atom:link href="https://pages.htgaa.org/2026a/beyza-cennet-batir/projects/individual-final-project/beyza_dehydrin_cryoprotection/index.xml" rel="self" type="application/rss+xml"/></channel></rss>