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Johannes Köster, Myles Brown, X Shirley Liu
MOTIVATION: Multiplexed error-robust fluorescence in-situ hybridization (MERFISH) is a recent technology to obtain spatially resolved gene or transcript expression profiles in single cells for hundreds to thousands of genes in parallel. So far, no statistical framework to analyze MERFISH data is available. RESULTS: We present a Bayesian model for single cell transcript expression analysis on MERFISH data. We show that the model successfully captures uncertainty in MERFISH data and eliminates systematic biases that can occur in raw RNA molecule counts obtained with MERFISH...
March 15, 2019: Bioinformatics
Shahar Barbash, Torbjörn Persson, Emily Lorenzen, Manija A Kazmi, Thomas Huber, Thomas P Sakmar
A recent phylogenetic analysis showed global co-evolution of G protein-coupled receptors (GPCRs) and receptor-activity-modifying proteins (RAMPs) suggesting global interactions between these two protein families. Experimental validation of these findings is challenging because in humans whereas there are only three genes encoding RAMPs, there are about 800 genes encoding GPCRs. Here, we report an experimental approach to evaluate GPCR-RAMP interactions. As a proof-of-concept experiment, we over-expressed RAMP2 in HEK293T cells and evaluated the effect on the transcriptional levels of 14 representative GPCRs that were selected based on the earlier phylogenetic analysis...
December 31, 2018: IScience
Johannes Köster, Myles Brown, X Shirley Liu
No abstract text is available yet for this article.
October 25, 2018: Bioinformatics
Guiping Wang, Jeffrey R Moffitt, Xiaowei Zhuang
A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has not been fixed in the paper.
April 19, 2018: Scientific Reports
Guiping Wang, Jeffrey R Moffitt, Xiaowei Zhuang
As an image-based single-cell transcriptomics approach, multiplexed error-robust fluorescence in situ hybridization (MERFISH) allows hundreds to thousands of RNA species to be identified, counted and localized in individual cells while preserving the native spatial context of RNAs. In MERFISH, RNAs are identified via a combinatorial labeling approach that encodes RNA species with error-robust barcodes followed by sequential rounds of single-molecule FISH (smFISH) to read out these barcodes. The accuracy of RNA identification relies on spatially separated signals from individual RNA molecules, which limits the density of RNAs that can be measured and makes the multiplexed imaging of a large number of high-abundance RNAs challenging...
March 19, 2018: Scientific Reports
Jeffrey R Moffitt, Junjie Hao, Guiping Wang, Kok Hao Chen, Hazen P Babcock, Xiaowei Zhuang
Image-based approaches to single-cell transcriptomics, in which RNA species are identified and counted in situ via imaging, have emerged as a powerful complement to single-cell methods based on RNA sequencing of dissociated cells. These image-based approaches naturally preserve the native spatial context of RNAs within a cell and the organization of cells within tissue, which are important for addressing many biological questions. However, the throughput of these image-based approaches is relatively low. Here we report advances that lead to a drastic increase in the measurement throughput of multiplexed error-robust fluorescence in situ hybridization (MERFISH), an image-based approach to single-cell transcriptomics...
September 27, 2016: Proceedings of the National Academy of Sciences of the United States of America
J R Moffitt, X Zhuang
Quantitative measurements of both the copy number and spatial distribution of large fractions of the transcriptome in single cells could revolutionize our understanding of a variety of cellular and tissue behaviors in both healthy and diseased states. Single-molecule fluorescence in situ hybridization (smFISH)-an approach where individual RNAs are labeled with fluorescent probes and imaged in their native cellular and tissue context-provides both the copy number and spatial context of RNAs but has been limited in the number of RNA species that can be measured simultaneously...
2016: Methods in Enzymology
Alex K Shalek, Rahul Satija
A new paper in Science by Zhuang and colleagues introduces multiplexed error-robust FISH (MERFISH). MERFISH extends single-molecule imaging techniques to profile the copy number and localization patterns of thousands of genes, representing a major advance for spatial transcriptomics, with exciting potential applications in immunology.
July 2015: Trends in Immunology
Kok Hao Chen, Alistair N Boettiger, Jeffrey R Moffitt, Siyuan Wang, Xiaowei Zhuang
Knowledge of the expression profile and spatial landscape of the transcriptome in individual cells is essential for understanding the rich repertoire of cellular behaviors. Here, we report multiplexed error-robust fluorescence in situ hybridization (MERFISH), a single-molecule imaging approach that allows the copy numbers and spatial localizations of thousands of RNA species to be determined in single cells. Using error-robust encoding schemes to combat single-molecule labeling and detection errors, we demonstrated the imaging of 100 to 1000 distinct RNA species in hundreds of individual cells...
April 24, 2015: Science
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