New tools for highly accessible single cell RNA sequencing

项目总结/摘要 单细胞RNA测序（scRNAseq）提供了前所未有的洞察基因调控途径， 复杂组织中的活细胞目前的单细胞技术可以提供高质量、高分辨率的 分析细胞群体中的扰动，但在访问、可负担性和可扩展性方面受到限制 细胞输入。为了广泛地部署scRNAseq作为高通量、成本有效的研究和发现， 作为一种工具，平台需要几个特性：检测基因表达中细微变化的灵敏度， 药物干预的结果，可扩展性，以便有效地处理数百到数百个 数千个输入单元，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。PIPseq 为scRNAseq提供了前所未有的可访问性，适用于不易通过 竞争技术。消除对复杂仪器和昂贵微流体的依赖 耗材允许即使是中等装备的细胞生物学实验室实施强大的scRNAseq测定。的 PIPseq平台的固有可扩展性在许多患者样本 或者可以评估治疗条件。这一直接进入第二阶段的建议将建立 通过严格的内部稳定性和再现性测试， 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 评估由学术合作者提供的原发性乳腺组织样本。

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.