New tools for highly accessible single cell RNA sequencing

项目摘要/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。目前的单电池技术可以提供高质量、高分辨率 分析蜂窝种群中的扰动，但在可获得性、可负担性和可扩展性方面受到限制 单元格输入的。为了将scRNAseq作为高通量、高成本效益的研究和发现广泛部署 工具，平台需要几个特征：敏感度检测基因表达的细微变化 药物干预的结果，可伸缩性，以便高效地处理数百到数百个 数千个输入信元，能够处理多种控制和处理条件的协议，以及 高效的多路复用排序功能，实现了成本效益。 Fluent BioSciences开发了用于单细胞RNA测序的预模板化即时分区 (PIPseq)作为一种新的scRNAseq方法，与现有方法相比具有关键优势。PIPSeq 为应用程序提供前所未有的scRNAseq可访问性 相互竞争的技术。消除对复杂仪器和昂贵的微流体的依赖 消耗品允许即使是装备简陋的细胞生物学实验室也可以进行强大的scRNAseq分析。这个 PIPseq平台的内在可伸缩性在许多患者样本的应用中也特别有用 或者可以对治疗条件进行评估。这项直接转到第二阶段的提案将建立 通过严格的内部稳定性和重复性测试，PIPseq试剂盒的商业就绪状态，外部 与多个基因组学核心实验室合作者以及临床前进行评估和重复性测试 由学术合作者提供的原始乳腺组织样本的评估。

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.