Cited by 1,101Single-cell and spatial transcriptomicsGenomics and Phylogenetic StudiesInnovative Microfluidic and Catalytic Techniques Innovation

The DOI matches its Crossref record; 59 of the 60 references it cites were found in a registry or an index; 22 of the 30 sources its claims cite were checked for support.

Multiplexed droplet single-cell RNA-sequencing using natural genetic variation

Open access

Published version of a preprint

Abstract

Droplet single-cell RNA-sequencing (dscRNA-seq) has enabled rapid, massively parallel profiling of transcriptomes. However, assessing differential expression across multiple individuals has been hampered by inefficient sample processing and technical batch effects. Here we describe a computational tool, demuxlet, that harnesses natural genetic variation to determine the sample identity of each droplet containing a single cell (singlet) and detect droplets containing two cells (doublets). These capabilities enable multiplexed dscRNA-seq experiments in which cells from unrelated individuals are pooled and captured at higher throughput than in standard workflows. Using simulated data, we show that 50 single-nucleotide polymorphisms (SNPs) per cell are sufficient to assign 97% of singlets and identify 92% of doublets in pools of up to 64 individuals. Given genotyping data for each of eight pooled samples, demuxlet correctly recovers the sample identity of >99% of singlets and identifies doublets at rates consistent with previous estimates. We apply demuxlet to assess cell-type-specific changes in gene expression in 8 pooled lupus patient samples treated with interferon (IFN)-β and perform eQTL analysis on 23 pooled samples.

Overview

Integrity signals

Registry

Crossref · 10.1038/nbt.4042

Cites

59 of 60 references verified0 citation errors · 1 not checkable

Statistics

Nothing to recompute.

Verification

Claims the paper makes

22 of 30 cited sources checked · 3 supported · 0 flagged · accuracy not yet measured

Show all 32 claims

  • Probably not a substantial claim. 11
  • It may state two separate claims as one. 28
35 of 35
  • The paper relies on this without support.

    In standard workflows, dscRNA-seq of many samples in parallel remains challenging to implement.

    “However, in standard workflows, dscRNA-seq of many samples in parallel remains challenging to implement.”
  • The authors state this as an assumption.

    When a base call is incorrect, the probability of the observed base call given the genotype and error follows the distribution in Supplementary Table 4.

    “When ecvi = 1, we assume that Pr(bcvi|g, ecvi) follows Supplementary Table 4.”
  • The authors state this as an assumption.

    The latent variable indicating whether a base call is correct follows a Bernoulli distribution with parameter 10−qcvi10.

    “ecvi is assumed to follow Bernoulli (10−qcvi10) where qcvi is a phred-scale quality score of the observed base call.”
  • The paper relies on this without support.

    The prior probability of genotype g follows a Binomial(2, pv) distribution, where pv is the population allele frequency of the alternate allele.

    “Pr(g) ~ Binomial(2, pv) and pv is the population allele frequency of the alternate allele.”
  • The authors state this as an assumption.

    The expected rate of multiplets for a given number of loaded cells follows an exponential model: d(x)=1−(1−d0)^(x/x0).

    “Then the expected multiplet rates when x cells are loaded can be modeled exponentially as d(x)=1−(1−d0)xx0.”
5 of 5

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • W1 and W2 are assigned to different pools

    Minor

    The initial W1 and W2 wells contain four donors each, but the later passage calls the first two additional pools W1 and W2 even though they contain seven and eight donors. The text does not distinguish these as reused labels.

1 of 1

Citations

Cites

Flagged first, then by citations

Show all 59 references

Cited by

Sorted by citations

Items 601–700 of 998

Metadata

Authors
Kang, H. M., Subramaniam, M., Targ, S., Nguyen, M., Maliskova, L., McCarthy, E., Wan, E., Wong, S., Byrnes, L., Lanata, C. M., Gate, R. E., Mostafavi, S., Marson, A., Zaitlen, N., Criswell, L. A., & Ye, C. J.
Affiliations
  • University of Michigan (US)
  • University of California, San Francisco (US)
  • University of British Columbia (CA)
Corresponding author
  • Kang, Hyun Min
  • Ye, Chun Jimmie
Venue
Nature Biotechnology
Published
2017-12-11
Volume
36
Issue
1
Pages
89-94
ISSN
  • 0733-222X (ISSN-L)
  • 1087-0156
  • 1546-1696
OpenAlex
W2774307122
Cited by
1,101
References
59
Citations by year
2026: 163 · 2025: 142 · 2024: 182 · 2023: 134 · 2022: 116 · 2021: 153 · 2020: 105 · 2019: 65 · 2018: 37 · 2017: 3
FWCI
20.46
Type
journal article
Language
en
Open access
Locations
  • Nature Biotechnology (publishedVersion)
  • bioRxiv (Cold Spring Harbor Laboratory) (submittedVersion)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (acceptedVersion)
  • PubMed Central (acceptedVersion)
  • eScholarship (California Digital Library) (submittedVersion)
Funders
  • National Heart, Lung, and Blood Institute (K25HL121295, U01HL137182)
  • National Institute of General Medical Sciences (T32GM007618, T32GM067547)
  • National Institute of Allergy and Infectious Diseases (R38AI140299, U19AI090023, R21AI133337)
  • National Institute of Diabetes and Digestive and Kidney Diseases (F30DK115167)
  • National Institute of Arthritis and Musculoskeletal and Skin Diseases (R01AR071522, P30AR070155)
  • National Institute of Dental and Craniofacial Research (R03DE025665)
  • National Center for Chronic Disease Prevention and Health Promotion (U01 DP006486, U01 DP005120)
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
  • Physical Sciences › Chemical Engineering › Fluid Flow and Transfer Processes › Innovative Microfluidic and Catalytic Techniques Innovation
Keywords
natural genetic variation, single nucleotide polymorphisms, doublet detection, pooled samples, batch effects, cell type-specific gene expression, expression quantitative trait loci, interferon-beta, lupus
MeSH terms
Genotype, Humans, Interferons, Lupus Erythematosus, Systemic, Polymorphism, Single Nucleotide, Quantitative Trait Loci, Single-Cell Analysis, High-Throughput Nucleotide Sequencing, Transcriptome

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

A language model drew the claims from the paper on 2026-10-06, under version 3 of the claim definition (model deepseek-v4.1-flash, prompt extract-v3).

A second model labelled each claim on 2026-10-06 (model jev-1.13-20260917, definition revision gold-guideline-v0.2).

Cited sources are checked with a language model’s help (gpt-6-sol).

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review