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新技术实现固定组织中蛋白质和RNA的多重数字空间分析
2020-05-12 14:23

美国NanoString技术公司Joseph M. Beechem、Christopher R. Merritt等研究人员合作,开发出固定组织中蛋白质和RNA的多重数字空间分析技术。该项研究成果发表于2020年5月11日出的《自然—生物技术》。

研究人员研发的数字空间分析(DSP)是一种适用于福尔马林固定、石蜡包埋(FFPE)样品的蛋白质或RNA空间分析方法。该方法依赖于(1)使用寡核苷酸标签对蛋白质或RNA进行多重读取;(2)通过光切割(PC)接头与亲和试剂(抗体或RNA探针)连接的寡核苷酸标签;(3)剪切光投射到组织样品上,以PC寡核苷酸以任何空间模式释放覆盖1至5,000个细胞的目标区域(ROI)。
 
DSP能够使用抗体读数在ROI内实现单细胞敏感性,并且RNA检测可适用至约600个单独的mRNA转录本。研究人员通过使用nCounter系统显示了淋巴样、结直肠肿瘤和自身免疫组织中多达44种蛋白质和96个基因(928个RNA探针)的空间概况分析,并通过使用下一代测序(NGS)显示了1,412个基因(4,998个RNA探针)。DSP不仅可以用于分析生物库样品中的蛋白质和RNA,而且可以用于分析患者样品中的免疫标记物,具有影响临床决策的潜力。
 
附:英文原文

Title: Multiplex digital spatial profiling of proteins and RNA in fixed tissue

Author: Christopher R. Merritt, Giang T. Ong, Sarah E. Church, Kristi Barker, Patrick Danaher, Gary Geiss, Margaret Hoang, Jaemyeong Jung, Yan Liang, Jill McKay-Fleisch, Karen Nguyen, Zach Norgaard, Kristina Sorg, Isaac Sprague, Charles Warren, Sarah Warren, Philippa J. Webster, Zoey Zhou, Daniel R. Zollinger, Dwayne L. Dunaway, Gordon B. Mills, Joseph M. Beechem

Issue&Volume: 2020-05-11

Abstract: Digital Spatial Profiling (DSP) is a method for highly multiplex spatial profiling of proteins or RNAs suitable for use on formalin-fixed, paraffin-embedded (FFPE) samples. The approach relies on (1) multiplexed readout of proteins or RNAs using oligonucleotide tags; (2) oligonucleotide tags attached to affinity reagents (antibodies or RNA probes) through a photocleavable (PC) linker; and (3) photocleaving light projected onto the tissue sample to release PC oligonucleotides in any spatial pattern across a region of interest (ROI) covering 1 to ~5,000 cells. DSP is capable of single-cell sensitivity within an ROI using the antibody readout, with RNA detection feasible down to ~600 individual mRNA transcripts. We show spatial profiling of up to 44 proteins and 96 genes (928 RNA probes) in lymphoid, colorectal tumor and autoimmune tissues by using the nCounter system and 1,412 genes (4,998 RNA probes) by using next-generation sequencing (NGS). DSP may be used to profile not only proteins and RNAs in biobanked samples but also immune markers in patient samples, with potential prognostic and predictive potential for clinical decision-making.

DOI: 10.1038/s41587-020-0472-9

Source: https://www.nature.com/articles/s41587-020-0472-9

Nature Biotechnology:《自然—生物技术》,创刊于1996年。隶属于施普林格·自然出版集团,最新IF:68.164
官方网址:https://www.nature.com/nbt/
投稿链接:https://mts-nbt.nature.com/cgi-bin/main.plex


本期文章:《自然—生物技术》:Online/在线发表

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