Supplement to Advancing CRISPR-Cas Technologies for Discovery and Characterization of Novel Fungal Natural Products

先进 CRISPR-Cas 技术的补充，用于新型真菌天然产物的发现和表征

• 批准号: 10393788
• 负责人: Xue Gao
• 金额: $ 0.84万
• 依托单位: RICE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10393788
• 关键词: Academia; Anabolism; Bioinformatics; CRISPR/Cas technology; Characteristics; Chemical Structure; Chemicals; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Development; Drug Industry; Drug resistance; Enzymes; Fungal Genome; Gene Cluster; Genes; Genetic; Genome engineering; Goals; Human; Investigation; Medicine; Molds; Natural Products; Parents; Pathway interactions; Peptides; Pharmacologic Substance; Pharmacology; Play; Property; Research; Ribosomes; Role; Source; Technology; Text; Therapeutic; algorithm development; analog; base; bioinformatics tool; drug candidate; drug discovery; fungus; genetic manipulation; genome editing; human disease; interest; next generation; novel; novel therapeutics; tool

Project Summary/Abstract (30 lines of text) Fungal natural products (NPs) have been a preeminent source of medicine and played pivotal roles as pharmaceuticals for the treatment of human diseases. The rapid expansion of fungal genome sequences and the development of bioinformatics tools have enabled the identification of thousands of fungal NP biosynthetic gene clusters (BGCs), thus providing an unprecedented opportunity to discover new fungal NPs. However, the discovery of new bioactive fungal NPs remains challenging, due to difficulties in prioritizing BGCs and genetic manipulations in fungi. In this proposal, we expect to build pipelines to rapidly discover novel bioactive fungal natural products that can serve as the next generation of drug candidates for the treatment of human diseases; to do this, we will apply the CRISPR Cas genome editing technologies and dedicate these tools to the biosynthesis of fungal natural products. To achieve the research goal, our first direction will focus on identifying and characterizing rarely discovered ribosomally synthesized and post-translationally modified peptides (RiPPs) from fungal origins. Due to RiPPs’ unique biosynthetic machinery, complex chemical characteristics, and important pharmacological properties, bacterial RiPPs have drawn strong interest from both academia and the pharmaceutical industry. However, only a handful of RiPPs have been identified from fungi, even though fungi is known to be a prolific producer of NPs. By characterizing novel biosynthetic enzymes of known RiPPs and new fungal BGCs identified by bioinformatics analysis, we expect to greatly broaden and deepen our understanding of the biosynthesis of fungal RiPPs and expand the repertoire of novel fungal RiPP NPs. Our second direction will focus on expanding and applying CRISPR-based genome engineering toolkits to characterize biosynthetic gene clusters from filamentous fungi. CRISPR-Cas tools have been successfully demonstrated to be feasible in fungal species but are rarely applied in the investigation of fungal NP biosynthesis. We will develop complementary sets of CRISPR-Cas tools for manipulating fungal biosynthetic gene clusters in both native and heterologous expression hosts. By doing so, we expect to develop a full set of CRISPR gene-editing toolkits to rapidly carry out genetic manipulations to study natural product biosynthesis in filamentous fungi. Together, the two research directions and collaborative research endeavors through BGC characterization, genetic tool advancement, and new bioinformatics algorithm development will build a complete pipeline to significantly increase the repertoire of fungal NPs and analogs, especially fungal RiPPs, making these molecules valuable drug candidates for human therapeutics.

项目摘要/摘要（30 行文本） 真菌天然产物 (NP) 一直是卓越的药物来源，并发挥着关键作用 作为治疗人类疾病的药物的作用 真菌的迅速扩张。 基因组序列和生物信息学工具的发展使得鉴定成为可能 数千个真菌NP生物合成基因簇（BGC），从而提供了前所未有的 发现新的真菌纳米颗粒的机会 然而，新的生物活性真菌纳米颗粒的发现。 由于在真菌中优先考虑 BGC 和遗传操作仍然具有挑战性。 根据这项提案，我们期望建立管道来快速发现新型生物活性真菌天然物质 可作为治疗人类的下一代候选药物的产品 为此，我们将应用 CRISPR Cas 基因组编辑技术并致力于 这些工具能够生物合成真菌天然产物，以实现我们的第一个研究目标。 方向将侧重于识别和表征罕见发现的核糖体合成 以及源自真菌的翻译后修饰肽 (RiPP)，因为 RiPP 具有独特的特性。 生物合成机制、复杂的化学特性和重要的药理作用 细菌 RiPP 的特性引起了学术界和科学界的浓厚兴趣 然而，即使是在制药行业，也仅从真菌中鉴定出少数 RiPP。 尽管已知真菌是纳米粒子的多产者，但通过表征新型生物合成。 通过生物信息学分析鉴定出已知 RiPP 和新真菌 BGC 的酶，我们预计 大大拓宽和加深我们对真菌 RiPP 生物合成的理解，并扩展 我们的第二个方向将集中于扩展和开发新型真菌 RiPP NP。 应用基于 CRISPR 的基因组工程工具包来表征生物合成基因簇 来自丝状真菌的 CRISPR-Cas 工具已被成功证明是可行的。 在真菌物种中，但很少应用于真菌 NP 生物合成的研究。 开发用于操纵真菌生物合成基因的互补 CRISPR-Cas 工具集 通过这样做，我们期望开发一个在天然和异源表达宿主中的簇。 全套CRISPR基因编辑工具包，快速进行基因操作研究自然 丝状真菌中的产物生物合成是两个研究方向的共同点和协作。 通过 BGC 表征、遗传工具进步和新的研究努力 生物信息学算法开发将构建完整的管道，显着提高 真菌 NP 和类似物的全部成分，尤其是真菌 RiPP，使这些分子变得有价值 用于人类治疗的候选药物。