SBIR PHASE I TOPIC #439 - ADVANCED SAMPLE PROCESSING PLATFORMS FOR DOWNSTREAM SINGLE-CELL MULTI-OMIC ANALYSIS

SBIR 第一阶段主题

• 批准号: 10724107
• 负责人: QI ZHAO
• 金额: $ 39.98万
• 依托单位: ENRICH THERAPEUTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-19 至 2023-09-18
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10724107
• 关键词: Cell Line; Cell Separation; Cells; Digestion; Dissociation; Genomics; Goals; Hand; Human; Image; Individual; Libraries; Liver neoplasms; Manuals; Maps; Monitor; Mus; Phase; Protocols documentation; Reagent; Research Design; Resistance; Retrieval; Running; Sampling; Site; Small Business Innovation Research Grant; Time; Tissues; Tumor Tissue; Validation; base; human tissue; multiple omics; operation; pancreatic neoplasm; real-time images; tumor

The long term goal of this proposal is to develop a fast, hands-free platform that integrates tissue dissociation, clone characterization and isolation to support the growing field of single celll multiomics. During the phase I of the proposal, enrich will adapt the exiting image based, debris-resistant clone isolation machine by developing essential reagents, protocols and consumables for in-place tissue dissociation, multicriteria single cell isolation and lossless single cell retrieval. Efficiency of tissue digestion, single cell capture, washing and retrieval will be tightly monitored by existing real-time imaging functionality. Adequate and high viability single cells from human liver and pancreas tumors will be isolated and scRNA and scDNA sequencing libraries of each sample will be prepared by 10xGenomics platform and submitted for Illumina sequencing. The validation will be achieved by an array of metrics including total operation time, level of manual intevention, single cell yield and quality and sc sequence mappability. if successful, a multi-site, tumor type study will be performed in the next phase.

这项提议的长期目标是开发一种快速、免提的平台，集组织分离、克隆鉴定和分离于一体，以支持不断增长的单细胞多组学领域。在提议的第一阶段，Enrich将通过开发用于就地组织分离、多标准单细胞分离和无损单细胞检索的基本试剂、协议和消耗品，对现有的基于图像的、防碎片的克隆分离机进行改造。组织消化、单细胞捕获、洗涤和回收的效率将通过现有的实时成像功能进行密切监测。将从人肝和胰腺肿瘤中分离出足够和高活性的单个细胞，并利用10x基因组学平台制备每个样本的scRNA和scDNA测序文库，并提交Illumina测序。验证将通过一系列指标实现，包括总操作时间、人工干预水平、单细胞产量和质量以及sc序列可映射性。如果成功，下一阶段将进行多部位、肿瘤类型的研究。