烯二炔类化合物是迄今发现抗肿瘤活性最强的一类天然产物，目前仅报道15个(含4个已芳香环化的衍生物)化合物，却有两个已开发成药品和一个Ⅱ临床。虽然高通量测序和生物信息学技术的迅速发展，但把所有的菌株进行测序和注释来挖掘沉默基因簇资源仍然不切实际。基于高通量测序、生物信息学技术和前期研究基础，本项目选择一株具有价值潜力的烯二炔合成海洋放线菌161111作为优先菌株，以抗肿瘤活性为导向, 通过CRISPR/Cas9基因组编辑技术阻断或过表达该代谢途径中重要的结构基因、调控基因和抗性基因, 结合比较代谢图谱技术, 分离和鉴定该类天然产物, 并建立烯二炔化合物次级代谢产物资源的基因组挖掘技术。同时，阐述烯二炔类合成基因的相关功能，加深对烯二炔类合成机制的认识，为将来运用生物组合学技术获得更多具有临床应用价值的新型烯二炔类化合物奠定基础，从而创新烯二炔天然产物的发现策略。

The enediyne natural products are the most cytotoxic molecules in existence today, among the 15 enediynes (with an additional four isolated in the cycloaromatized form) known to date, two, neocarzinostatin (NCS) and calicheamicin (CAL), have been developed into clinical drugs and one, C-1027, is in clinical trials Ⅱ. In spite of the rapid advances in DNA sequencing technologies and bioinformatics, it is still impractical to sequence and annotate all strains as a practical means to discover silent natural product biosynthetic gene clusters. Based on high-throughput sequencing, bioinformatics and our preliminary research foundation, the strain Jishengella sp. 161111 could then be devoted preferentially to interrogate strains that hold the highest promise in producing novel enediyne natural products. To induce or enhance the expression of cryptic or poorly expressed pathway to provied material for struction elucidation and biological testing, bioactivity-guided strategy is to find ways that blocking or over-expression of the metabolic pathways of important structural genes, regulatory genes and resistance gene, based of the CRISPR/Cas9 toolkitfor genome editing in Jishengella sp. 161111. The enediyne natural product will be isolated and identified by comparative metabolic profiling of mutant and wild-type strains. Furthermore, the described genome mining approach may offer an effective strategy to mine for the novel metabolic products of silent natural product biosynthetic gene clusters in verrucosisporae. Understanding of the regulation and functional assignments of these ORFs in enediyne biosynthetic gene clusters, and characterizing the enediyne biosynthetic machinery, new enediynes are bound to be discovered using combinatorial biosynthesis approaches, therefore holds a great promise for anticancer drug discovery. The study could fundamentally change how enediyne natural products are discovered.