Activation of Fungal Silent Gene Clusters

With the rapid development of high-throughput sequencing technology and bioinformatics, maximizing the activation of silent gene clusters has become the key to the efficient utilization of fungal resources in the post-genomic era. Although many silent gene clusters exist in fungi, whether these gene clusters are simply in a silent state or have undergone genetic mutations to lose their functions needs further analysis. We aim to improve the discrimination between silent and inactivated gene clusters and develop repair techniques for inactivated genes based on this, thus improving the activation efficiency of silent gene clusters. Lifeasible can provide a variety of non-directed activation and directed activation strategies to achieve the expression of fungal silencing gene clusters. On this basis, a large number of natural compounds with novel structures and outstanding activities can be obtained.

Figure 1. Strategies for transcriptional activation for fungal biosynthetic gene clusters. (Mózsik L, et al., 2022)

• One strain of many compounds for activation of silent fungal gene clusters. The simplest and most direct way to activate silent gene clusters is to use different media to ferment the target strains, i.e., the OSMAC strategy. We offer this service to help our customers explore the secondary metabolites produced by the target strains in different media.
• Co-cultivation for activation of silent fungal gene clusters. There are complex interactions between microorganisms that can induce the synthesis of structurally diverse metabolites from each other. We can help our customers screen for suitable activation conditions based on microbial co-culture strategies to improve the activation efficiency of silenced gene clusters.
• Epigenetic regulation-mediated activation of silent gene clusters. In fungi, epigenetic modifications are closely related to synthesizing secondary metabolites. For example, DNA methylation has known effects on gene transcription, and acetylation of histones usually activates gene expression. We can improve the activation efficiency of silenced gene clusters by adding epigenetic small molecule inhibitors or knocking out epigenetically regulated genes, among other strategies.
• Global Regulator-Based Activation of Silent Gene Clusters. Global regulators regulate the expression of multiple genes simultaneously under specific induction conditions. By utilizing global transcription factors, we can help our customers achieve targeted activation of silent gene clusters.
• Activation of silent gene clusters mediated by promoter substitution. Promoter replacement is using a new promoter to replace the original promoter of a silent gene, thus allowing the silent gene to be expressed normally in the original host. We can use a promoter replacement strategy to achieve targeted activation of silent gene clusters.
• Heterologous expression-mediated activation of silent gene clusters. Heterologous expression is a strategy for targeted activation of silent gene clusters, and we can provide Saccharomyces cerevisiae and Aspergillus spp. as heterologous expression hosts for activation of silent gene clusters in genetically manipulated strains where promoter substitution strategies are not applicable.
• CRISPR/Cas genome editing in plant fungi.CRISPR/Cas genome editing technologies include gene knockout, knock-in, and point mutations that can modify the genome sequence more precisely. We can use CRISPR/Cas technology to activate unknown gene clusters in beneficial fungi to discover new secondary metabolites.
• Construction of high enzyme-producing fungal strains. We use genetic modification technologies to help customers build highly enzyme-producing fungal strains by improving the nutrient sensing, gene translation, protein transcription, and secretion capabilities of fungi.

Lifeasible specializes in fungal silent gene cluster activation and offers a variety of strategies to help customers maximize the activation of fungal silent gene clusters. As your trusted partner, we can meet all your fungal phylogenetic analysis needs and provide you with efficient and high-quality services. If you want to know the details, please contact us.