Advanced Single Cell Technology Innovation Core

项目摘要 拟议中的癌症系统治疗（CastT）中心专注于了解和靶向分子 不同但共存的细胞状态和转变，建立和维持肿瘤和肿瘤细胞的生长。 系统层面的微环境。这些目标需要具有单细胞分辨率的分析方法。的 先进的单细胞技术创新核心（ASCeTIC）将提供最先进的技术， 细胞基因组分析，多重单细胞分子扰动，并在原位验证，以卡斯特研究人员。 我们将提供一套工具，包括用于联合单细胞RNA/ATAC-seq、单细胞RNA/ATAC-seq和单细胞RNA/ATAC-seq的多组学方法。 成像/RNA-seq（SCOPE-seq）和单细胞CRISPRi/RNA-seq（CROP-seq）。确认必须 用于原位验证和肿瘤微环境中细胞-细胞相互作用研究的空间分布， 我们还将提供免疫组织化学分析，RNA-FISH和循环免疫荧光（cycIF）， 同时定量单个组织切片中单个细胞中的数十个蛋白质表位。

PROJECT SUMMARY The proposed Cancer Systems Therapeutics (CaST) Center focuses on understanding and targeting molecularly distinct yet co-existing cell states and transitions that establish and maintain tumors and tumor microenvironments at a systems level. These goals demand analytical methods with single-cell resolution. The Advanced Single Cell Technology Innovation Core (ASCeTIC) will provide state-of-the-art technology for single- cell genomic analysis, multiplexed single-cell molecular perturbation, and in situ validation to CaST investigators. We will offer a suite of tools including multi-omic methods for joint single-cell RNA/ATAC-seq, single-cell imaging/RNA-seq (SCOPE-seq), and single-cell CRISPRi/RNA-seq (CROP-seq). Recognizing the importance of spatial profiling for both in situ validation and studies of cell-cell interactions in the tumor microenvironment, we will also offer immunohistochemical analysis, RNA-FISH, and cyclic immunofluorescence (cycIF), which can simultaneously quantify tens of protein epitopes in individual cells in a single tissue section.