Ensemblex: an accuracy-weighted ensemble genetic demultiplexing framework for population-scale scRNAseq sample pooling

Abstract Multiplexing samples from distinct individuals prior to sequencing is a promising step towards achieving population-scale single-cell RNA sequencing by reducing the restrictive costs of the technology. Individual genetic demultiplexing tools resolve the donor-of-origin identity of pooled ce...

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Main Authors: Michael R. Fiorini, Saeid Amiri, Allison A. Dilliott, Cristine M. Yde Ohki, Lukasz Smigielski, Susanne Walitza, Edward A. Fon, Edna Grünblatt, Rhalena A. Thomas, Sali M. K. Farhan
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Language:English
Published: BMC 2025-07-01
Series:Genome Biology
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Online Access:https://doi.org/10.1186/s13059-025-03643-1
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author Michael R. Fiorini
Saeid Amiri
Allison A. Dilliott
Cristine M. Yde Ohki
Lukasz Smigielski
Susanne Walitza
Edward A. Fon
Edna Grünblatt
Rhalena A. Thomas
Sali M. K. Farhan
author_facet Michael R. Fiorini
Saeid Amiri
Allison A. Dilliott
Cristine M. Yde Ohki
Lukasz Smigielski
Susanne Walitza
Edward A. Fon
Edna Grünblatt
Rhalena A. Thomas
Sali M. K. Farhan
author_sort Michael R. Fiorini
collection DOAJ
description Abstract Multiplexing samples from distinct individuals prior to sequencing is a promising step towards achieving population-scale single-cell RNA sequencing by reducing the restrictive costs of the technology. Individual genetic demultiplexing tools resolve the donor-of-origin identity of pooled cells using natural genetic variation but present diminished accuracy on highly multiplexed experiments, impeding the analytic potential of the dataset. In response, we introduce Ensemblex: an accuracy-weighted, ensemble genetic demultiplexing framework that integrates four distinct algorithms to identify the most probable subject labels. Using computationally and experimentally pooled samples, we demonstrate Ensemblex’s superior accuracy and illustrate the implications of robust demultiplexing on biological analyses.
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institution Kabale University
issn 1474-760X
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publishDate 2025-07-01
publisher BMC
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spelling doaj-art-c19bbee6db9649c3b9029639c11d8d6e2025-08-20T04:01:36ZengBMCGenome Biology1474-760X2025-07-0126113910.1186/s13059-025-03643-1Ensemblex: an accuracy-weighted ensemble genetic demultiplexing framework for population-scale scRNAseq sample poolingMichael R. Fiorini0Saeid Amiri1Allison A. Dilliott2Cristine M. Yde Ohki3Lukasz Smigielski4Susanne Walitza5Edward A. Fon6Edna Grünblatt7Rhalena A. Thomas8Sali M. K. Farhan9Department of Human Genetics, McGill UniversityThe Montreal Neurological Institute-Hospital, McGill UniversityThe Montreal Neurological Institute-Hospital, McGill UniversityDepartment of Child and Adolescent Psychiatry and Psychotherapy, Psychiatric University Hospital of Zurich, University of ZurichDepartment of Child and Adolescent Psychiatry and Psychotherapy, Psychiatric University Hospital of Zurich, University of ZurichDepartment of Child and Adolescent Psychiatry and Psychotherapy, Psychiatric University Hospital of Zurich, University of ZurichThe Montreal Neurological Institute-Hospital, McGill UniversityDepartment of Child and Adolescent Psychiatry and Psychotherapy, Psychiatric University Hospital of Zurich, University of ZurichThe Montreal Neurological Institute-Hospital, McGill UniversityDepartment of Human Genetics, McGill UniversityAbstract Multiplexing samples from distinct individuals prior to sequencing is a promising step towards achieving population-scale single-cell RNA sequencing by reducing the restrictive costs of the technology. Individual genetic demultiplexing tools resolve the donor-of-origin identity of pooled cells using natural genetic variation but present diminished accuracy on highly multiplexed experiments, impeding the analytic potential of the dataset. In response, we introduce Ensemblex: an accuracy-weighted, ensemble genetic demultiplexing framework that integrates four distinct algorithms to identify the most probable subject labels. Using computationally and experimentally pooled samples, we demonstrate Ensemblex’s superior accuracy and illustrate the implications of robust demultiplexing on biological analyses.https://doi.org/10.1186/s13059-025-03643-1Single-cell RNA sequencingMultiplexingSample poolingGenetic demultiplexingInduced pluripotent stem cellsDifferential gene expression
spellingShingle Michael R. Fiorini
Saeid Amiri
Allison A. Dilliott
Cristine M. Yde Ohki
Lukasz Smigielski
Susanne Walitza
Edward A. Fon
Edna Grünblatt
Rhalena A. Thomas
Sali M. K. Farhan
Ensemblex: an accuracy-weighted ensemble genetic demultiplexing framework for population-scale scRNAseq sample pooling
Genome Biology
Single-cell RNA sequencing
Multiplexing
Sample pooling
Genetic demultiplexing
Induced pluripotent stem cells
Differential gene expression
title Ensemblex: an accuracy-weighted ensemble genetic demultiplexing framework for population-scale scRNAseq sample pooling
title_full Ensemblex: an accuracy-weighted ensemble genetic demultiplexing framework for population-scale scRNAseq sample pooling
title_fullStr Ensemblex: an accuracy-weighted ensemble genetic demultiplexing framework for population-scale scRNAseq sample pooling
title_full_unstemmed Ensemblex: an accuracy-weighted ensemble genetic demultiplexing framework for population-scale scRNAseq sample pooling
title_short Ensemblex: an accuracy-weighted ensemble genetic demultiplexing framework for population-scale scRNAseq sample pooling
title_sort ensemblex an accuracy weighted ensemble genetic demultiplexing framework for population scale scrnaseq sample pooling
topic Single-cell RNA sequencing
Multiplexing
Sample pooling
Genetic demultiplexing
Induced pluripotent stem cells
Differential gene expression
url https://doi.org/10.1186/s13059-025-03643-1
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