New tools for highly accessible single cell RNA sequencing

易于使用的单细胞 RNA 测序新工具

基本信息

批准号：
10579927
负责人：
Kristina Fontanez
金额：
$ 77.52万
依托单位：
FLUENT BIOSCIENCES INC.
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-03-01 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10579927
关键词：
Acceleration Address Adopted Aging Bar Codes Biological Assay Biological Sciences Breast Capital Cell Separation Cells Cellular biology Characteristics Clinical Clinical Research Collaborations Complementary DNA Complex Consumption Contracts Cytolysis DNA Sequencing Facility Data Dependence Estrogens Evaluation Experimental Designs Feedback Gene Expression Generations Genes Genetic Transcription Genomics Human Immunology Individual Industry Informatics Intervention Laboratories Libraries Life Malignant Neoplasms Mammary Gland Parenchyma Marketing Methods Microfluidics Molecular Biology Patients Performance Pharmacologic Substance Phase Physiological Population Population Heterogeneity Preparation Process Production Progesterone Protocols documentation Readiness Reagent Regulator Genes Regulatory Pathway Reproducibility Research Research Personnel Resected Resolution Resource Sharing Sampling Site Small Business Innovation Research Grant Specific qualifier value System Technology Testing Time Tissue Sample Tissues Validation Variant cell preparation cell type clinical investigation clinically relevant commercialization cost cost effective cost effectiveness flexibility indexing insight instrumentation manufacture neuron development novel strategies preclinical evaluation recruit sequencing platform single cell technology single-cell RNA sequencing tool transcriptomics usability validation studies wasting web portal

项目摘要

PROJECT SUMMARY / ABSTRACT Single cell RNA sequencing (scRNAseq) provides unprecedented insight into the gene regulatory pathways of living cells in complex tissues. Current single cell technologies can provide high-quality, high resolution analysis of perturbations in cellular populations, but are limited in terms of access, affordability, and scalability of cell input. In order to broadly deploy scRNAseq as a high-throughput, cost effective research and discovery tool, a platform requires several characteristics: sensitivity to detect subtle changes in gene expression as a result of pharmaceutical intervention, scalability in order to efficiently process hundreds to hundreds of thousands of input cells, a protocol capable of processing multiple control and treatment conditions, and efficient multiplexed sequencing capability for cost-effectiveness. Fluent BioSciences has developed Pre-templated Instant Partitions for single cell RNA sequencing (PIPseq) as a novel approach toward scRNAseq that has key advantages over existing methods. PIPseq provides unprecedented accessibility to scRNAseq for applications that are not readily addressable by competing technologies. Eliminating the dependence on complex instrumentation and expensive microfluidic consumables allows even modestly equipped cell biology labs to implement powerful scRNAseq assays. The intrinsic scalability of the PIPseq platform is also particularly useful in applications where many patient samples or treatment conditions might be evaluated. This direct-to-phase II proposal will establish commercial-readiness of PIPseq kits through rigorous internal stability and reproducibility testing, external evaluation and reproducibility testing with multiple genomics core laboratory collaborators, and pre-clinical evaluation of primary breast tissue samples provided by an academic collaborator.

项目概要/摘要单细胞 RNA 测序 (scRNAseq) 提供了对基因调控途径的前所未有的洞察复杂组织中的活细胞。当前的单细胞技术可以提供高质量、高分辨率分析细胞群体的扰动，但在访问、负担能力和可扩展性方面受到限制的单元格输入。为了广泛部署 scRNAseq 作为高通量、具有成本效益的研究和发现工具，一个平台需要几个特性：检测基因表达细微变化的敏感性药物干预的结果，可扩展性，以便有效地处理数百到数百个数千个输入细胞，能够处理多种控制和治疗条件的协议，以及高效的多重测序能力，具有成本效益。 Fluent BioSciences 开发了用于单细胞 RNA 测序的预模板即时分区（PIPseq）作为 scRNAseq 的一种新方法，与现有方法相比具有关键优势。 PIP序列为 scRNAseq 无法轻易解决的应用程序提供前所未有的可访问性竞争技术。消除对复杂仪器和昂贵的微流体的依赖即使是装备一般的细胞生物学实验室也可以使用耗材来实施强大的 scRNAseq 检测。这 PIPseq 平台的内在可扩展性在需要大量患者样本的应用中也特别有用或者可以评估治疗条件。这项直接进入第二阶段的提案将建立通过严格的内部稳定性和再现性测试、外部测试，PIPseq 试剂盒的商业准备就绪与多个基因组学核心实验室合作者进行评估和再现性测试，以及临床前对学术合作者提供的初级乳腺组织样本进行评估。