Commercialization of methods and reagents for high-throughput single cell analysis

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Fluent BioSciences开发了预模板即时分区， 单细胞RNA测序（PIPseq），消除了定制微流体的成本和复杂性， 该仪器具有简单、灵活的工作流程，可在任何实验室环境中实施。 通过第一阶段SBIR支持（1 R43 GM 137648 - 01 A1），我们已经证明PIPseq本质上是 可扩展的，并且能够在单个反应中捕获数百至数百万个细胞。新事物的出现， 高容量和低成本的测序模式，例如由Ultima Genomics开发的测序模式， 细胞输入单细胞转录组学不仅可行，而且实用， 实验模式在本提案中，我们将扩大PIPseq单细胞分析的能力， 为包括深层组织在内的高细胞输入应用提供简单、可扩展且具有成本效益的解决方案 分析、多重药物测试和合并的CRISPR筛选。

PROJECT SUMMARY / ABSTRACT Single-cell RNA sequencing (scRNAseq) provides unprecedented insight into the gene regulatory pathways of living cells in complex tissues. Fluent BioSciences has developed Pretemplated Instant Partitioning for single-cell RNA sequencing (PIPseq) that eliminates the cost and complexity of bespoke microfluidics and instrumentation and features a simple, flexible workflow that can be implemented in any laboratory setting. With Phase I SBIR support (1 R43 GM137648-01A1) we have demonstrated that PIPseq is intrinsically scalable and is capable of capturing from hundreds to millions of cells in a single reaction. The advent of new, high capacity, and low cost sequencing modalities such as that developed by Ultima Genomics makes high cell-input single cell transcriptomics not only feasible but practical, enabling previously inaccessible experimental modalities. In this proposal, we will expand the capabilities of PIPseq single cell analysis to provide simple, scalable, and cost-effective solutions for high cell input applications including deep tissue profiling, multiplexed drug testing, and pooled CRISPR screening.