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Long-read whole-genome analysis of human single cells
Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm, Sweden; Department of Biosystems Science and Engineering, ETH Zurich, Basel, Switzerland; ETH AI Center, ETH Zurich, Zurich, Switzerland.
Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm, Sweden.
Department of Molecular Medicine and Surgery, Karolinska Institutet, Stockholm, Sweden; Department of Clinical Genetics, Karolinska University Hospital, Stockholm, Sweden.
Science for Life Laboratory, Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden.
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2023 (English)In: Nature Communications, E-ISSN 2041-1723, Vol. 14, no 1, article id 5164Article in journal (Refereed) Published
Abstract [en]

Long-read sequencing has dramatically increased our understanding of human genome variation. Here, we demonstrate that long-read technology can give new insights into the genomic architecture of individual cells. Clonally expanded CD8+ T-cells from a human donor were subjected to droplet-based multiple displacement amplification (dMDA) to generate long molecules with reduced bias. PacBio sequencing generated up to 40% genome coverage per single-cell, enabling detection of single nucleotide variants (SNVs), structural variants (SVs), and tandem repeats, also in regions inaccessible by short reads. 28 somatic SNVs were detected, including one case of mitochondrial heteroplasmy. 5473 high-confidence SVs/cell were discovered, a sixteen-fold increase compared to Illumina-based results from clonally related cells. Single-cell de novo assembly generated a genome size of up to 598 Mb and 1762 (12.8%) complete gene models. In summary, our work shows the promise of long-read sequencing toward characterization of the full spectrum of genetic variation in single cells.

Place, publisher, year, edition, pages
Springer Nature , 2023. Vol. 14, no 1, article id 5164
National Category
Medical Genetics and Genomics Genetics and Genomics Bioinformatics (Computational Biology)
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URN: urn:nbn:se:kth:diva-336707DOI: 10.1038/s41467-023-40898-3ISI: 001188258200002PubMedID: 37620373Scopus ID: 2-s2.0-85168664713OAI: oai:DiVA.org:kth-336707DiVA, id: diva2:1798198
Note

QC 20240402

Available from: 2023-09-18 Created: 2023-09-18 Last updated: 2025-02-10Bibliographically approved

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