Genetic Storage-Expansive Dualistic Strategies for Engineering Cryo-resistant Bioartificial Tissues – A Meta-Review

Abstract

Cryogenic bioartificial tissue preservation remains a critical challenge, primarily due to the reliance on cytotoxic cryoprotectants for ice crystal inhibition. Recent advances in synthetic biology offer promising alternatives by enabling the endogenous intracellular cryoprotection. This review explores the potential of genetically-encoded cryoresistance, drawing insights from natural systems such as antifreeze proteins (AFPs), heat-shock proteins (HSPs), and cold-adaptive pathways. It reviews key molecular features—solvation thermodynamics, hydrogen-bonding networks, and membrane-stabilizing motifs—used to inhibit ice formation and mitigate damage in engineered tissues. Computational and experimental studies indicate that genetic modifications promoting the expression of cryoactive proteins can enhance cell viability across thermal transitions, from normothermic culture to cryogenic storage. Furthermore, we discuss how synthetic gene circuits and pathways can optimize cryoresistance while minimizing metabolic and structural disintegration. By integrating biophysical modelling with genetic design principles, this review highlights strategies for developing next-generation bioartificial tissues capable of even, ubiquitous, scalable and endogenous cold adaptation. Finally, we assess translational challenges, including scalability, safety, and regulatory considerations, while outlining future directions for clinical implementation. This genetic engineering paradigm holds significant potential to overcome the limitations of traditional cryopreservation, advancing the field towards more robust and clinically viable tissue-based therapies.