Computational and analytical challenges in single-cell transcriptomics
Abstract
The development of high-throughput RNA sequencing (RNA-seq) at the single-cell level has already led to profound new discoveries in biology, ranging from the identification of novel cell types to the study of global patterns of stochastic gene expression. Alongside the technological breakthroughs that have facilitated the large-scale generation of single-cell transcriptomic data, it is important to consider the specific computational and analytical challenges that still have to be overcome. Although some tools for analysing RNA-seq data from bulk cell populations can be readily applied to single-cell RNA-seq data, many new computational strategies are required to fully exploit this data type and to enable a comprehensive yet detailed study of gene expression at the single-cell level.
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Single-cell RNA-seq data exhibits significant cell-to-cell variation due to technical factors, including the number of molecules detected in each cell, which can confound biological heterogeneity with technical effects.
“cell-to-cell variation in technical factors can also confound these results [1, 2].”
From Normalization and variance stabilization of single-cell RNA-seq data using regularized negative binomial regression (2019) · cites this paper as [2]
Multiple replicates of C1 collections from the same biological condition are necessary to facilitate estimation of technical variability even with ERCC spike-in controls.
“Consequently, multiple replicates of C1 collections from the same biological condition are necessary to facilitate estimation of technical variability even with the presence of ERCC spike-in controls10.”
From Batch effects and the effective design of single-cell gene expression studies (2017) · cites this paper as [10]
ERCCs can be problematic for modeling endogenous mRNAs, potentially due to their shorter length, shorter poly-A tail, and missing 5' cap.
“It has been noted before that ERCCs can be problematic for modeling endogenous mRNAs (Risso et al., 2014), potentially due to their shorter length, shorter poly-A tail, and their missing 5 0 cap (Gr € un and van Oudenaarden, 2015; Stegle et al., 2015).”
From Comparative Analysis of Single-Cell RNA Sequencing Methods (2017) · cites this paper as Stegle et al., 2015
Extra Poisson variability includes biological variation and technical variation, and technical variation includes noise introduced by amplification.
“This extra Poisson variability includes biological variation (assumed to be the same across methods in our data) and technical variation, and the latter includes noise introduced by amplification (Brennecke et al., 2013; Gr € un et al., 2014; Stegle et al., 2015).”
From Comparative Analysis of Single-Cell RNA Sequencing Methods (2017) · cites this paper as Stegle et al., 2015
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SCANPY: large-scale single-cell gene expression data analysis
Wolf, F. A., Angerer, P., & Theis, F. J. (2018) · Genome Biology · doi:10.1186/s13059-017-1382-0 · Cited by 8,881 · see citing works
Scenic: single-cell regulatory network inference and clustering.
Aibar, S., González-Blas, C. B., Moerman, T., Huynh-Thu, V. A., Imrichova, H., Hulselmans, G., Rambow, F., Marine, J.-C., Geurts, P., Aerts, J., et al. (2017) · Nature Methods · doi:10.1038/nmeth.4463 · Cited by 6,520 · see citing works
McGinnis, C. S., Murrow, L. M., & Gartner, Z. J. (2019) · Cell Systems · doi:10.1016/j.cels.2019.03.003 · Cited by 4,521 · see citing works
Hafemeister, C., & Satija, R. (2019) · Genome Biology · doi:10.1186/s13059-019-1874-1 · Cited by 4,479 · see citing works
“While unsupervised analysis of single-cell data has transformative potential to uncover heterogeneous cell types and states, cell-to-cell variation in technical factors can also confound these results [1, 2].”
BackgroundDetermining cell type abundance and expression from bulk tissues with digital cytometry
Newman, A. M., Steen, C. B., Liu, C. L., Gentles, A. J., Chaudhuri, A. A., Scherer, F., Khodadoust, M. S., Esfahani, M. S., Luca, B. A., Steiner, D. F., et al. (2019) · Nature Biotechnology · doi:10.1038/s41587-019-0114-2 · Cited by 4,427 · see citing works
A survey of best practices for RNA-seq data analysis
Conesa, A., Madrigal, P., Tarazona, S., Gomez-Cabrero, D., Cervera, A., McPherson, A., Szcześniak, M. W., Gaffney, D. J., Elo, L. L., Zhang, X., et al. (2016) · Genome Biology · doi:10.1186/s13059-016-0881-8 · Cited by 2,794 · see citing works
Regev, A., Teichmann, S. A., Lander, E. S., Amit, I., Benoist, C., Birney, E., Bodenmiller, B., Campbell, P., Carninci, P., Clatworthy, M., et al. (2017) · eLife · doi:10.7554/elife.27041 · Cited by 2,232 · see citing works
Self-citationRNA sequencing: the teenage years
Stark, R., Grzelak, M., & Hadfield, J. (2019) · Nature Reviews Genetics · doi:10.1038/s41576-019-0150-2 · Cited by 1,906 · see citing works
Recovering Gene Interactions from Single-Cell Data Using Data Diffusion
van Dijk, D., Sharma, R., Nainys, J., Yim, K., Kathail, P., Carr, A. J., Burdziak, C., Moon, K. R., Chaffer, C. L., Pattabiraman, D., et al. (2018) · Cell · doi:10.1016/j.cell.2018.05.061 · Cited by 1,829 · see citing works
Wolf, F. A., Hamey, F. K., Plass, M., Solana, J., Dahlin, J. S., Göttgens, B., Rajewsky, N., Simon, L., & Theis, F. J. (2019) · Genome Biology · doi:10.1186/s13059-019-1663-x · Cited by 1,698 · see citing works
Single-Cell Transcriptome Profiling of Human Pancreatic Islets in Health and Type 2 Diabetes
Segerstolpe, Å., Palasantza, A., Eliasson, P., Andersson, E.-M., Andréasson, A.-C., Sun, X., Picelli, S., Sabirsh, A., Clausen, M., Bjursell, M. K., et al. (2016) · Cell Metabolism · doi:10.1016/j.cmet.2016.08.020 · Cited by 1,550 · see citing works
A step-by-step workflow for low-level analysis of single-cell RNA-seq data
Lun, A. T. L., McCarthy, D. J., & Marioni, J. C. (2016) · F1000Research · doi:10.12688/f1000research.9501.1 · Cited by 1,511 · see citing works
Self-citationComparative Analysis of Single-Cell RNA Sequencing Methods
Ziegenhain, C., Vieth, B., Parekh, S., Reinius, B., Guillaumet-Adkins, A., Smets, M., Leonhardt, H., Heyn, H., Hellmann, I., & Enard, W. (2017) · Molecular Cell · doi:10.1016/j.molcel.2017.01.023 · Cited by 1,494 · see citing works
The Technology and Biology of Single-Cell RNA Sequencing
Kolodziejczyk, A. A., Kim, J. K., Svensson, V., Marioni, J. C., & Teichmann, S. A. (2015) · Molecular Cell · doi:10.1016/j.molcel.2015.04.005 · Cited by 1,487 · see citing works
Self-citationSimultaneous enumeration of cancer and immune cell types from bulk tumor gene expression data
Racle, J., de Jonge, K., Baumgaertner, P., Speiser, D. E., & Gfeller, D. (2017) · eLife · doi:10.7554/elife.26476 · Cited by 1,373 · see citing works
Multiplexed droplet single-cell RNA-sequencing using natural genetic variation
Kang, H. M., Subramaniam, M., Targ, S., Nguyen, M., Maliskova, L., McCarthy, E., Wan, E., Wong, S., Byrnes, L., Lanata, C. M., et al. (2018) · Nature Biotechnology · doi:10.1038/nbt.4042 · Cited by 1,339 · see citing works
“Improvements in biochemistry 11, 12 and microfluidics 13, 14 continue to increase the number of cells and transcripts profiled per experiment.”
BackgroundEleven grand challenges in single-cell data science
Lähnemann, D., Köster, J., Szczurek, E., McCarthy, D. J., Hicks, S. C., Robinson, M. D., Vallejos, C. A., Campbell, K. R., Beerenwinkel, N., Mahfouz, A., et al. (2020) · Genome Biology · doi:10.1186/s13059-020-1926-6 · Cited by 1,327 · see citing works
Self-citationChallenges in unsupervised clustering of single-cell RNA-seq data
Kiselev, V. Y., Andrews, T. S., & Hemberg, M. (2019) · Nature Reviews Genetics · doi:10.1038/s41576-018-0088-9 · Cited by 1,149 · see citing works
Landscape of X chromosome inactivation across human tissues
GTEx Consortium, Tukiainen, T., Villani, A.-C., Yen, A., Rivas, M. A., Marshall, J. L., Satija, R., Aguirre, M., Gauthier, L., Fleharty, M., et al. (2017) · Nature · doi:10.1038/nature24265 · Cited by 1,138 · see citing works
Buettner, F., Natarajan, K. N., Casale, F. P., Proserpio, V., Scialdone, A., Theis, F. J., Teichmann, S. A., Marioni, J. C., & Stegle, O. (2015) · Nature Biotechnology · doi:10.1038/nbt.3102 · Cited by 1,136 · see citing works
Self-citationPooling across cells to normalize single-cell RNA sequencing data with many zero counts
L. Lun, A. T., Bach, K., & Marioni, J. C. (2016) · Genome Biology · doi:10.1186/s13059-016-0947-7 · Cited by 1,108 · see citing works
Self-citationGene co-expression analysis for functional classification and gene–disease predictions
van Dam, S., Võsa, U., van der Graaf, A., Franke, L., & de Magalhães, J. P. (2017) · Briefings in Bioinformatics · doi:10.1093/bib/bbw139 · Cited by 1,086 · see citing works
Single-cell RNA-seq denoising using a deep count autoencoder
Eraslan, G., Simon, L. M., Mircea, M., Mueller, N. S., & Theis, F. J. (2019) · Nature Communications · doi:10.1038/s41467-018-07931-2 · Cited by 1,084 · see citing works
A practical guide to single-cell RNA-sequencing for biomedical research and clinical applications
Haque, A., Engel, J., Teichmann, S. A., & Lönnberg, T. (2017) · Genome Medicine · doi:10.1186/s13073-017-0467-4 · Cited by 1,071 · see citing works
Self-citationClustering trees: a visualization for evaluating clusterings at multiple resolutions
Zappia, L., & Oshlack, A. (2018) · GigaScience · doi:10.1093/gigascience/giy083 · Cited by 1,064 · see citing works
Integrating single-cell and spatial transcriptomics to elucidate intercellular tissue dynamics
Longo, S. K., Guo, M. G., Ji, A. L., & Khavari, P. A. (2021) · Nature Reviews Genetics · doi:10.1038/s41576-021-00370-8 · Cited by 1,059 · see citing works
Stoeckius, M., Zheng, S., Houck-Loomis, B., Hao, S., Yeung, B. Z., Mauck, W. M., Smibert, P., & Satija, R. (2018) · Genome Biology · doi:10.1186/s13059-018-1603-1 · Cited by 1,046 · see citing works
Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)
Cossarizza, A., Chang, H., Radbruch, A., Acs, A., Adam, D., Adam‐Klages, S., Agace, W. W., Aghaeepour, N., Akdiş, M., Allez, M., et al. (2019) · European Journal of Immunology · doi:10.1002/eji.201970107 · Cited by 982 · see citing works
Single-Cell RNA-Seq Technologies and Related Computational Data Analysis
Chen, G., Ning, B., & Shi, T. (2019) · Frontiers in Genetics · doi:10.3389/fgene.2019.00317 · Cited by 913 · see citing works
Efficient integration of heterogeneous single-cell transcriptomes using Scanorama
Hie, B., Bryson, B. D., & Berger, B. (2019) · Nature Biotechnology · doi:10.1038/s41587-019-0113-3 · Cited by 853 · see citing works
Defining cell types and states with single-cell genomics
Trapnell, C. (2015) · Genome Research · doi:10.1101/gr.190595.115 · Cited by 813 · see citing works
Single-Cell RNA-Seq with Waterfall Reveals Molecular Cascades underlying Adult Neurogenesis
Shin, J., Berg, D. A., Zhu, Y., Shin, J. Y., Song, J., Bonaguidi, M. A., Enikolopov, G., Nauen, D. W., Christian, K. M., Ming, G.-L., et al. (2015) · Cell Stem Cell · doi:10.1016/j.stem.2015.07.013 · Cited by 784 · see citing works
Dissecting Immune Circuits by Linking CRISPR-Pooled Screens with Single-Cell RNA-Seq
Jaitin, D. A., Weiner, A., Yofe, I., Lara-Astiaso, D., Keren-Shaul, H., David, E., Salame, T. M., Tanay, A., van Oudenaarden, A., & Amit, I. (2016) · Cell · doi:10.1016/j.cell.2016.11.039 · Cited by 777 · see citing works
The origin and evolution of cell types
Arendt, D., Musser, J. M., Baker, C. V. H., Bergman, A., Cepko, C., Erwin, D. H., Pavlicev, M., Schlosser, G., Widder, S., Laubichler, M. D., et al. (2016) · Nature Reviews Genetics · doi:10.1038/nrg.2016.127 · Cited by 764 · see citing works
RNA ‐Seq methods for transcriptome analysis
Hrdličková, R., Toloue, M., & Tian, B. (2016) · Wiley Interdisciplinary Reviews - RNA · doi:10.1002/wrna.1364 · Cited by 686 · see citing works
Zitnik, M., Nguyen, F., Wang, B., Leskovec, J., Goldenberg, A., & Hoffman, M. M. (2019) · Information Fusion · doi:10.1016/j.inffus.2018.09.012 · Cited by 675 · see citing works
MULTI-seq: sample multiplexing for single-cell RNA sequencing using lipid-tagged indices
McGinnis, C. S., Patterson, D. M., Winkler, J., Conrad, D. N., Hein, M. Y., Srivastava, V., Hu, J. L., Murrow, L. M., Weissman, J. S., Werb, Z., et al. (2019) · Nature Methods · doi:10.1038/s41592-019-0433-8 · Cited by 643 · see citing works
<i>destiny</i> : diffusion maps for large-scale single-cell data in R
Angerer, P., Haghverdi, L., Büttner, M., Theis, F. J., Marr, C., & Buettner, F. (2016) · Bioinformatics · doi:10.1093/bioinformatics/btv715 · Cited by 640 · see citing works
Missing data and technical variability in single-cell RNA-sequencing experiments
Hicks, S. C., Townes, F. W., Teng, M., & Irizarry, R. A. (2018) · Biostatistics · doi:10.1093/biostatistics/kxx053 · Cited by 635 · see citing works
Comparison and evaluation of statistical error models for scRNA-seq
Choudhary, S., & Satija, R. (2022) · Genome Biology · doi:10.1186/s13059-021-02584-9 · Cited by 634 · see citing works
Revealing the vectors of cellular identity with single-cell genomics
Wagner, A., Regev, A., & Yosef, N. (2016) · Nature Biotechnology · doi:10.1038/nbt.3711 · Cited by 632 · see citing works
Single-cell RNA sequencing for the study of development, physiology and disease
Potter, S. S. (2018) · Nature Reviews Nephrology · doi:10.1038/s41581-018-0021-7 · Cited by 621 · see citing works
Gene Regulatory Network Inference from Single-Cell Data Using Multivariate Information Measures
Chan, T. E., Stumpf, M. P. H., & Babtie, A. C. (2017) · Cell Systems · doi:10.1016/j.cels.2017.08.014 · Cited by 578 · see citing works
Single Cell RNA-Sequencing of Pluripotent States Unlocks Modular Transcriptional Variation
Kolodziejczyk, A. A., Kim, J. K., Tsang, J. C. H., Ilicic, T., Henriksson, J., Natarajan, K. N., Tuck, A. C., Gao, X., Bühler, M., Liu, P., et al. (2015) · Cell Stem Cell · doi:10.1016/j.stem.2015.09.011 · Cited by 570 · see citing works
Self-citationPower analysis of single-cell RNA-sequencing experiments
Svensson, V., Natarajan, K. N., Ly, L.-H., Miragaia, R. J., Labalette, C., Macaulay, I. C., Cvejic, A., & Teichmann, S. A. (2017) · Nature Methods · doi:10.1038/nmeth.4220 · Cited by 568 · see citing works
Self-citationWishbone identifies bifurcating developmental trajectories from single-cell data
Setty, M., Tadmor, M. D., Reich-Zeliger, S., Angel, O., Salame, T. M., Kathail, P., Choi, K., Bendall, S., Friedman, N., & Pe'er, D. (2016) · Nature Biotechnology · doi:10.1038/nbt.3569 · Cited by 558 · see citing works
Crowell, H. L., Soneson, C., Germain, P.-L., Calini, D., Collin, L., Raposo, C., Malhotra, D., & Robinson, M. D. (2020) · Nature Communications · doi:10.1038/s41467-020-19894-4 · Cited by 515 · see citing works
A test metric for assessing single-cell RNA-seq batch correction
Büttner, M., Miao, Z., Wolf, F. A., Teichmann, S. A., & Theis, F. J. (2019) · Nature Methods · doi:10.1038/s41592-018-0254-1 · Cited by 511 · see citing works
Self-citationCTCF and Cohesin in Genome Folding and Transcriptional Gene Regulation
Merkenschlager, M., & Nora, E. P. (2016) · Annual Review of Genomics and Human Genetics · doi:10.1146/annurev-genom-083115-022339 · Cited by 505 · see citing works
Heterogeneity in Oct4 and Sox2 Targets Biases Cell Fate in 4-Cell Mouse Embryos
Goolam, M., Scialdone, A., Graham, S. J. L., Macaulay, I. C., Jedrusik, A., Hupalowska, A., Voet, T., Marioni, J. C., & Zernicka-Goetz, M. (2016) · Cell · doi:10.1016/j.cell.2016.01.047 · Cited by 484 · see citing works
Self-citationChu, L.-F., Leng, N., Zhang, J., Hou, Z., Mamott, D., Vereide, D. T., Choi, J., Kendziorski, C., Stewart, R., & Thomson, J. A. (2016) · Genome Biology · doi:10.1186/s13059-016-1033-x · Cited by 477 · see citing works
scPred: accurate supervised method for cell-type classification from single-cell RNA-seq data
Alquicira-Hernández, J., Sathe, A., Ji, H. P., Nguyen, Q. H., & Powell, J. E. (2019) · Genome biology · doi:10.1186/s13059-019-1862-5 · Cited by 474 · see citing works
Computational principles and challenges in single-cell data integration
Argelaguet, R., Cuomo, A. S. E., Stegle, O., & Marioni, J. C. (2021) · Nature Biotechnology · doi:10.1038/s41587-021-00895-7 · Cited by 472 · see citing works
Self-citationSingle-cell multiomics: multiple measurements from single cells
Macaulay, I. C., Ponting, C. P., & Voet, T. (2017) · Trends in Genetics · doi:10.1016/j.tig.2016.12.003 · Cited by 451 · see citing works
Transition states and cell fate decisions in epigenetic landscapes
Moris, N., Pina, C., & Arias, A. M. (2016) · Nature Reviews Genetics · doi:10.1038/nrg.2016.98 · Cited by 438 · see citing works
Direct cell reprogramming: approaches, mechanisms and progress
Wang, H., Yang, Y., Liu, J., & Li, Q. (2021) · Nature Reviews Molecular Cell Biology · doi:10.1038/s41580-021-00335-z · Cited by 438 · see citing works
Design and computational analysis of single-cell RNA-sequencing experiments
Bacher, R. L., & Kendziorski, C. M. (2016) · Genome biology · doi:10.1186/s13059-016-0927-y · Cited by 437 · see citing works
Normalizing single-cell RNA sequencing data: challenges and opportunities
Vallejos, C. A., Risso, D., Scialdone, A., Dudoit, S., & Marioni, J. C. (2017) · Nature Methods · doi:10.1038/nmeth.4292 · Cited by 436 · see citing works
Self-citationSingle-cell RNA sequencing: Technical advancements and biological applications
Hedlund, E., & Deng, Q. (2017) · Molecular Aspects of Medicine · doi:10.1016/j.mam.2017.07.003 · Cited by 396 · see citing works
High-throughput spatial mapping of single-cell RNA-seq data to tissue of origin
Achim, K., Pettit, J.-B., Saraiva, L. R., Gavriouchkina, D., Larsson, T., Arendt, D., & Marioni, J. C. (2015) · Nature Biotechnology · doi:10.1038/nbt.3209 · Cited by 395 · see citing works
Self-citationComputational assignment of cell-cycle stage from single-cell transcriptome data
Scialdone, A., Natarajan, K. N., Saraiva, L. R., Proserpio, V., Teichmann, S. A., Stegle, O., Marioni, J. C., & Buettner, F. (2015) · Methods · doi:10.1016/j.ymeth.2015.06.021 · Cited by 386 · see citing works
Self-citationThe promise of spatial transcriptomics for neuroscience in the era of molecular cell typing
Lein, E., Borm, L. E., & Linnarsson, S. (2017) · Science · doi:10.1126/science.aan6827 · Cited by 383 · see citing works
Unravelling biology and shifting paradigms in cancer with single-cell sequencing
Baslan, T., & Hicks, J. B. (2017) · Nature reviews. Cancer · doi:10.1038/nrc.2017.58 · Cited by 377 · see citing works
The first five years of single-cell cancer genomics and beyondPreprint
Navin, N. E. (2015) · Genome Research · doi:10.1101/gr.191098.115 · Cited by 368 · see citing works
Single cell transcriptomics comes of age
Aldridge, S., & Teichmann, S. A. (2020) · Nature Communications · doi:10.1038/s41467-020-18158-5 · Cited by 360 · see citing works
Self-citationBatch effects and the effective design of single-cell gene expression studies
Tung, P.-Y., Blischak, J. D., Hsiao, C. J., Knowles, D. A., Burnett, J. E., Pritchard, J. K., & Gilad, Y. (2017) · Scientific Reports · doi:10.1038/srep39921 · Cited by 356 · see citing works
Single‐cell analysis reveals cancer stem cell heterogeneity in hepatocellular carcinoma
Zheng, H., Pomyen, Y., Hernandez, M. O., Li, C., Livák, F., Tang, W., Dang, H. T., Greten, T. F., Davis, J. L., Zhao, Y., et al. (2018) · Hepatology · doi:10.1002/hep.29778 · Cited by 350 · see citing works
Single-cell entropy for accurate estimation of differentiation potency from a cell’s transcriptome
Teschendorff, A. E., & Enver, T. (2017) · Nature Communications · doi:10.1038/ncomms15599 · Cited by 346 · see citing works
Beyond bulk: a review of single cell transcriptomics methodologies and applications
Kulkarni, A., Anderson, A. G., Merullo, D. P., & Konopka, G. (2019) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2019.03.001 · Cited by 344 · see citing works
Aging increases cell-to-cell transcriptional variability upon immune stimulation
Martinez-Jimenez, C. P., Eling, N., Chen, H.-C., Vallejos, C. A., Kolodziejczyk, A. A., Connor, F., Stojic, L., Rayner, T. F., Stubbington, M. J. T., Teichmann, S. A., et al. (2017) · Science · doi:10.1126/science.aah4115 · Cited by 339 · see citing works
Self-citationInterpretable dimensionality reduction of single cell transcriptome data with deep generative models
Ding, J., Condon, A., & Shah, S. P. (2018) · Nature Communications · doi:10.1038/s41467-018-04368-5 · Cited by 337 · see citing works
Holland, C. H., Tanevski, J., Perales-Patón, J., Gleixner, J., Kumar, M. P., Mereu, E., Joughin, B. A., Stegle, O., Lauffenburger, D. A., Heyn, H., et al. (2020) · Genome Biology · doi:10.1186/s13059-020-1949-z · Cited by 323 · see citing works
Self-citationDirect Comparative Analyses of 10X Genomics Chromium and Smart-Seq2
Wang, X., He, Y., Zhang, Q., Ren, X., & Zhang, Z. (2021) · Genomics Proteomics & Bioinformatics · doi:10.1016/j.gpb.2020.02.005 · Cited by 316 · see citing works
DrImpute: imputing dropout events in single cell RNA sequencing data
Gong, W., Kwak, I., Pota, P., Koyano‐Nakagawa, N., & Garry, D. J. (2018) · BMC Bioinformatics · doi:10.1186/s12859-018-2226-y · Cited by 311 · see citing works
Single-cell epigenomics: powerful new methods for understanding gene regulation and cell identity
Clark, S. J., Lee, H. J., Smallwood, S. A., Kelsey, G., & Reik, W. (2016) · Genome Biology · doi:10.1186/s13059-016-0944-x · Cited by 310 · see citing works
scds: computational annotation of doublets in single-cell RNA sequencing data
Bais, A. S., & Kostka, D. (2020) · Bioinformatics · doi:10.1093/bioinformatics/btz698 · Cited by 308 · see citing works
Single-cell RNA sequencing in cardiovascular development, disease and medicine
Paik, D. T., Cho, S., Tian, L., Chang, H. Y., & Wu, J. C. (2020) · Nature Reviews Cardiology · doi:10.1038/s41569-020-0359-y · Cited by 306 · see citing works
Exploring the single-cell RNA-seq analysis landscape with the scRNA-tools database
Zappia, L., Phipson, B., & Oshlack, A. (2018) · PLoS Computational Biology · doi:10.1371/journal.pcbi.1006245 · Cited by 304 · see citing works
BASiCS: Bayesian Analysis of Single-Cell Sequencing Data
Vallejos, C. A., Marioni, J. C., & Richardson, S. (2015) · PLOS Computational Biology · doi:10.1371/journal.pcbi.1004333 · Cited by 302 · see citing works
Self-citationpcaReduce: hierarchical clustering of single cell transcriptional profiles
Žurauskienė, J., & Yau, C. (2016) · BMC Bioinformatics · doi:10.1186/s12859-016-0984-y · Cited by 298 · see citing works
GiniClust: detecting rare cell types from single-cell gene expression data with Gini index
Jiang, L., Chen, H., Pinello, L., & Yuan, G. (2016) · Genome biology · doi:10.1186/s13059-016-1010-4 · Cited by 296 · see citing works
Resolving early mesoderm diversification through single-cell expression profiling
Scialdone, A., Tanaka, Y., Jawaid, W., Moignard, V., Wilson, N. K., Macaulay, I. C., Marioni, J. C., & Göttgens, B. (2016) · Nature · doi:10.1038/nature18633 · Cited by 293 · see citing works
Self-citationDisentangling neural cell diversity using single-cell transcriptomics
Poulin, J.-F., Tasic, B., Hjerling-Leffler, J., Trimarchi, J. M., & Awatramani, R. (2016) · Nature Neuroscience · doi:10.1038/nn.4366 · Cited by 293 · see citing works
Keren-Shaul, H., Kenigsberg, E., Jaitin, D. A., David, E., Paul, F., Tanay, A., & Amit, I. (2019) · Nature Protocols · doi:10.1038/s41596-019-0164-4 · Cited by 292 · see citing works
Transcriptome Profiling in Human Diseases: New Advances and Perspectives
Casamassimi, A., Federico, A., Rienzo, M., Esposito, S., & Ciccodicola, A. (2017) · International Journal of Molecular Sciences · doi:10.3390/ijms18081652 · Cited by 288 · see citing works
Wang, T., Li, B., Nelson, C. E., & Nabavi, S. (2019) · BMC Bioinformatics · doi:10.1186/s12859-019-2599-6 · Cited by 286 · see citing works
Pavličev, M., Wagner, G. P., Chavan, A. R., Owens, K. J., Maziarz, J. D., Dunn-Fletcher, C., Kallapur, S. G., Muglia, L. J., & Jones, H. N. (2017) · Genome Research · doi:10.1101/gr.207597.116 · Cited by 281 · see citing works
Machine learning and statistical methods for clustering single-cell RNA-sequencing data
Petegrosso, R., Li, Z., & Kuang, R. (2019) · Briefings in Bioinformatics · doi:10.1093/bib/bbz063 · Cited by 277 · see citing works
Machine Learning in Genomic Medicine: A Review of Computational Problems and Data Sets
Leung, M. K. K., Delong, A., Alipanahi, B., & Frey, B. J. (2015) · Proceedings of the IEEE · doi:10.1109/jproc.2015.2494198 · Cited by 276 · see citing works
CHETAH: a selective, hierarchical cell type identification method for single-cell RNA sequencing
de Kanter, J. K., Lijnzaad, P., Candelli, T., Margaritis, T., & Holstege, F. C. P. (2019) · Nucleic Acids Research · doi:10.1093/nar/gkz543 · Cited by 274 · see citing works
The CRUK IMAXT Grand Challenge Team, O’Flanagan, C. H., Campbell, K. R., Zhang, A. W., Kabeer, F., Lim, J. L., Biele, J., Eirew, P. D., Lai, D., McPherson, A., et al. (2019) · Genome biology · doi:10.1186/s13059-019-1830-0 · Cited by 265 · see citing works
Single-Cell RNA Sequencing Analysis: A Step-by-Step Overview
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- Authors
- Stegle, O., Teichmann, S. A., & Marioni, J. C.
- Affiliations
- European Bioinformatics Institute (GB)
- Corresponding author
- Marioni, John C.
- Venue
- Nature Reviews Genetics
- Published
- 2015-01-28
- Volume
- 16
- Issue
- 3
- Pages
- 133-145
- ISSN
- 1471-0056 (ISSN-L)
- 1471-0064
- OpenAlex
- W1988391018
- Cited by
- 1,170
- References
- 90
- Citations by year
- 2026: 40 · 2025: 83 · 2024: 85 · 2023: 100 · 2022: 106 · 2021: 122 · 2020: 130 · 2019: 132 · 2018: 112 · 2017: 120 · 2016: 102 · 2015: 35 · 2014: 1
- FWCI
- 11.92
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Reviews Genetics (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Single-cell and spatial transcriptomics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Phylogenetic Studies
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gene expression and cancer classification
- Keywords
- single-cell RNA sequencing, single-cell transcriptomics, computational challenges, data analysis, stochastic gene expression, cell type annotation, gene expression analysis
- MeSH terms
- Animals, Cell Differentiation, Eukaryotic Cells, Humans, Quality Control, RNA, Messenger, Software, Gene Expression Profiling, Gene Regulatory Networks, Single-Cell Analysis, High-Throughput Nucleotide Sequencing, Transcriptome
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