Development Core

项目摘要 单细胞组学的突破为研究各种生物提供了前所未有的机会 问题包括组织异质性、细胞活动动力学、命运决定和细胞对 压力。自2009年首次被描述以来，单细胞技术发展迅速，目前大多数 实验室正在使用10X基因组公司的设备进行单细胞RNA测序(scRNA-seq) 基于液滴微流体学。虽然功能强大，但这种方法成本较高且吞吐量相对较低。一个 单细胞技术的另一个缺点是由于组织分离而丢失空间信息， 因此，它们利基中的生理背景细胞是未知的。了解细胞中基因的表达 组织结构和形态的背景是理解细胞功能的关键，这是 尤其是在慢性肾脏疾病(CKD)中，损伤、修复和纤维化发生在 肾小管间质壁龛。SCORE核心是一个资源开发核心，将解决这些限制 通过开发和实施尖端技术以及所需的计算管道 解释结果数据，以支持生物医学资源核心和广泛的肾脏研究社区 全国范围内。SCORE CORE的目标是开发新的协议和代码，以阐明 CKD的病理生物学，始于基于裂解池条形码的单核多组学和高分辨率 基于原位测序的空间分辨转录组学。我们将开发协议以同时使用 高通量ATAC和RNA表达与测序(Share-Seq)，一种高度可扩展的方法 同一单个细胞中染色质可及性和基因表达的测量，以及计算 用于分析结果数据的管道。此外，SCORE CORE将为HIGH开发协议 基于Xum平台的空间转录分辨率原位测序。这些协议和 代码将高度支持我们中心的用户，以及广泛的联盟。Score Core还将 与国家奥布莱恩联盟和指导委员会互动，确定未来发展的优先事项 尖端技术随处可得。

Project Summary Breakthroughs in single-cell omics are providing unprecedented opportunities to investigate diverse biological questions including tissue heterogeneity, cell activity dynamics, fate determination and cellular responses to stress. Since first being described in 2009, single-cell technologies have developed rapidly, and currently most laboratories are performing single-cell RNA sequencing (scRNA-seq) using equipment from 10X Genomics based on droplet microfluidics. While powerful, this approach is expensive and relatively low throughput. An additional drawback to single cell technologies is that they lose spatial information due to tissue dissociation, so the physiological context cells in their niche is unknown. Understanding cellular gene expression in the context of tissue organization and morphology is critical to understanding cellular function and this is particularly true in chronic kidney disease (CKD) where injury, repair and fibrosis initiation occurs in the tubulointerstitial niche. The SCORE Core is a Resource Development Core that will address these limitations by developing and operationalizing cutting-edge technologies, and the computational pipelines required to interpret resulting data, to support the Biomedical Resource Cores and the broad kidney research community nationally. The aim of the SCORE Core is to develop new protocols and code that will illuminate the pathobiology of CKD, beginning with split pool barcoding based single nuclei multi-omics and high resolution in situ sequencing-based spatially resolved transcriptomics. We will develop protocols to employ simultaneous high-throughput ATAC and RNA expression with sequencing (SHARE-seq), a highly scalable approach for measurement of chromatin accessibility and gene expression in the same single cell, and the computational pipelines to analyze the resulting data. In addition, the SCORE Core will develop the protocols for high resolution in situ-sequencing based spatial transcriptomics using the Xenium platform. These protocols and code will be highly enabling to users in our Center, and to the Consortium broadly. The SCORE Core will also interact with the National O’Brien Consortium and Steering Committee to prioritize the development of future cutting edge technologies as they become available.