Single-cell Rapid Capture Hybridization sequencing reliably detects isoform usage and coding mutations in targeted genes Article Swipe
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· 2025
· Open Access
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· DOI: https://doi.org/10.1101/gr.279322.124
Single-cell long-read sequencing has transformed our understanding of isoform usage and the mutation heterogeneity between cells. Despite unbiased in-depth analysis, the low sequencing throughput often results in insufficient read coverage, thereby limiting our ability to perform mutation calling for specific genes. Here, we developed a single-cell Rapid Capture Hybridization sequencing (scRaCH-seq) method that demonstrates high specificity and efficiency in capturing targeted transcripts using long-read sequencing, allowing an in-depth analysis of mutation status and transcript usage for genes of interest. The method includes creating a probe panel for transcript capture, using barcoded primers for pooling and efficient sequencing via Oxford Nanopore Technologies platforms. scRaCH-seq is applicable to stored and indexed single-cell cDNA, which allows analysis to be combined with existing short-read RNA-seq data sets. In our investigation of BTK and SF3B1 genes in samples from patients with chronic lymphocytic leukemia (CLL), we detect SF3B1 isoforms and mutations with high sensitivity. Integration with short-read single-cell RNA sequencing (scRNA-seq) data reveals significant gene expression differences in SF3B1 -mutated CLL cells, although it does not impact the sensitivity of the anticancer drug venetoclax. scRaCH-seq's capability to study long-read transcripts of multiple genes makes it a powerful tool for single-cell genomics.
Related Topics
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1101/gr.279322.124
- https://genome.cshlp.org/content/early/2025/01/10/gr.279322.124.full.pdf
- OA Status
- hybrid
- Cited By
- 5
- References
- 41
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4406263596
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- OpenAlex ID
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https://openalex.org/W4406263596Canonical identifier for this work in OpenAlex
- DOI
-
https://doi.org/10.1101/gr.279322.124Digital Object Identifier
- Title
-
Single-cell Rapid Capture Hybridization sequencing reliably detects isoform usage and coding mutations in targeted genesWork title
- Type
-
articleOpenAlex work type
- Language
-
enPrimary language
- Publication year
-
2025Year of publication
- Publication date
-
2025-01-10Full publication date if available
- Authors
-
Hongke Peng, Jafar S. Jabbari, Luyi Tian, Changqing Wang, Yupei You, Chong Chyn Chua, Natasha S. Anstee, Noorul Amin, Andrew H. Wei, N. Davidson, Andrew W. Roberts, David C.S. Huang, Matthew E. Ritchie, Rachel ThijssenList of authors in order
- Landing page
-
https://doi.org/10.1101/gr.279322.124Publisher landing page
- PDF URL
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https://genome.cshlp.org/content/early/2025/01/10/gr.279322.124.full.pdfDirect link to full text PDF
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YesWhether a free full text is available
- OA status
-
hybridOpen access status per OpenAlex
- OA URL
-
https://genome.cshlp.org/content/early/2025/01/10/gr.279322.124.full.pdfDirect OA link when available
- Concepts
-
Biology, Single cell sequencing, Gene, Genetics, Computational biology, DNA sequencing, Deep sequencing, Cancer genome sequencing, Genomics, Massive parallel sequencing, Complementary DNA, Single-cell analysis, Mutation, Genome, Exome sequencing, CellTop concepts (fields/topics) attached by OpenAlex
- Cited by
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5Total citation count in OpenAlex
- Citations by year (recent)
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2025: 5Per-year citation counts (last 5 years)
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41Number of works referenced by this work
- Related works (count)
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10Other works algorithmically related by OpenAlex
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