13 Living fungal mycelium with suppressed or abolished fruit-forming ability is a self-healing 14 substance particularly valuable biomaterial for further engineering and development in 15 applications such as monitoring/sensing environmental changes and secreting signals. The 16 ability to suppress fungal fruiting is also a useful tool for maintaining stability (e.g., shape, 17 form) of a mycelium-based biomaterial with ease and lower cost.18 The objective of this present study is to provide a biochemical solution to regulate the fruiting 19 body formation to replace heat killing of mycelium during production. We discovered that 20 GSK-3 activity directly correlates with the development of fruiting bodies in fungi, especially 21 mushroom forming fungi such as Coprinopsis cinerea. By regulating GSK-3 expression and 22 activity, one can control the fungal fruiting body development. 23 We successfully demonstrated that treatment of an inhibitor of GSK-3 kinase activity resulted 24 in acceleration in mycelium growth rate, absence of fruiting body and general decrease in 25 GSK-3 gene expression. Therefore, GSK-3 inhibitor is suggested to be included in the 26 mycelium cultivation recipes for regulating the growth of fungal mycelium and for inhibiting 27 the development of fruiting bodies. This is the first report of using a GSK-3 inhibitor, such as 28 lithium or any other GSK-3 inhibitor, to suppress or abolish fruiting body formation in living 29 fungal mycelium-based biomaterial. It also provides an innovative strategy for easy, reliable, 30 and low cost maintenance of biomaterial containing live fungal mycelium. 31 32 33 Introduction 34 Fungal mycelium-based biomaterials are fast emerging in recent years. Mycelium is the 35 vegetative structure of fungi, mainly composed of natural polymers. Mycelium-based 36 biomaterials have a wide range of applications due to their controlled and tunable properties 37 during growth, its self-assembly, self-healing, environmentally responsive, and biodegradable 38 nature. The dried mycelium has strength, durability, and many other beneficial qualities: it is 39 nontoxic, fire-resistant, mold resistant, water-resistant, and a great thermal insulator, amongst 40 other salient features(1-13). Under proper circumstances, mycelium of many mushroom-41 forming fungi will aggregate to form mushrooms, which are the fruiting body spreading 42 spores (14). Not only the fruiting bodies will cause conformational changes of the mycelium 43 based materials, but also the spores may cause allergy and infection in susceptible population.44 In current production procedures of the mycelium-based materials, whole material are heated 45 or treated with fungicide to kill the living cells, to stop the fruiting body formation(15). Such 46 rendered mycelium-based materials retain few of the benefits of living material. Therefore, 47 new approaches are needed for inhibiting fruiting body formation while keeping the 48 mycelium alive, in order to produce living mycelium-based materials of desirable qualiti...