A novel single-cell RNA-sequencing approach and its applicability connecting genotype to phenotype in ageing disease
Abstract Single cell multi-omics analysis has the potential to yield a comprehensive understanding of the cellular events that underlie the basis of human diseases. The cardinal feature to access this information is the technology used for single-cell isolation, barcoding, and sequencing. Most curre...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2022-03-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-07874-1 |
| _version_ | 1819236777725001728 |
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| author | Orr Shomroni Maren Sitte Julia Schmidt Sabnam Parbin Fabian Ludewig Gökhan Yigit Laura Cecilia Zelarayan Katrin Streckfuss-Bömeke Bernd Wollnik Gabriela Salinas |
| author_facet | Orr Shomroni Maren Sitte Julia Schmidt Sabnam Parbin Fabian Ludewig Gökhan Yigit Laura Cecilia Zelarayan Katrin Streckfuss-Bömeke Bernd Wollnik Gabriela Salinas |
| author_sort | Orr Shomroni |
| collection | DOAJ |
| description | Abstract Single cell multi-omics analysis has the potential to yield a comprehensive understanding of the cellular events that underlie the basis of human diseases. The cardinal feature to access this information is the technology used for single-cell isolation, barcoding, and sequencing. Most currently used single-cell RNA-sequencing platforms have limitations in several areas including cell selection, documentation and library chemistry. In this study, we describe a novel high-throughput, full-length, single-cell RNA-sequencing approach that combines the CellenONE isolation and sorting system with the ICELL8 processing instrument. This method offers substantial improvements in single cell selection, documentation and capturing rate. Moreover, it allows the use of flexible chemistry for library preparations and the analysis of living or fixed cells, whole cells independent of sizing and morphology, as well as of nuclei. We applied this method to dermal fibroblasts derived from six patients with different segmental progeria syndromes and defined phenotype associated pathway signatures with variant associated expression modifiers. These results validate the applicability of our method to highlight genotype-expression relationships for molecular phenotyping of individual cells derived from human patients. |
| first_indexed | 2024-12-23T13:09:50Z |
| format | Article |
| id | doaj.art-718a794a31d34de3b3335bcd025ec31d |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-23T13:09:50Z |
| publishDate | 2022-03-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-718a794a31d34de3b3335bcd025ec31d2022-12-21T17:45:46ZengNature PortfolioScientific Reports2045-23222022-03-0112111410.1038/s41598-022-07874-1A novel single-cell RNA-sequencing approach and its applicability connecting genotype to phenotype in ageing diseaseOrr Shomroni0Maren Sitte1Julia Schmidt2Sabnam Parbin3Fabian Ludewig4Gökhan Yigit5Laura Cecilia Zelarayan6Katrin Streckfuss-Bömeke7Bernd Wollnik8Gabriela Salinas9NGS- Core Unit for Integrative Genomics, Institute of Human Genetics, University Medical Center GöttingenNGS- Core Unit for Integrative Genomics, Institute of Human Genetics, University Medical Center GöttingenInstitute of Human Genetics, University Medical Center GöttingenNGS- Core Unit for Integrative Genomics, Institute of Human Genetics, University Medical Center GöttingenNGS- Core Unit for Integrative Genomics, Institute of Human Genetics, University Medical Center GöttingenInstitute of Human Genetics, University Medical Center GöttingenInstitute of Pharmacology and Toxicology, University Medical Center GöttingenClinic of Cardiology and Pneumology, University Medical Center GöttingenInstitute of Human Genetics, University Medical Center GöttingenNGS- Core Unit for Integrative Genomics, Institute of Human Genetics, University Medical Center GöttingenAbstract Single cell multi-omics analysis has the potential to yield a comprehensive understanding of the cellular events that underlie the basis of human diseases. The cardinal feature to access this information is the technology used for single-cell isolation, barcoding, and sequencing. Most currently used single-cell RNA-sequencing platforms have limitations in several areas including cell selection, documentation and library chemistry. In this study, we describe a novel high-throughput, full-length, single-cell RNA-sequencing approach that combines the CellenONE isolation and sorting system with the ICELL8 processing instrument. This method offers substantial improvements in single cell selection, documentation and capturing rate. Moreover, it allows the use of flexible chemistry for library preparations and the analysis of living or fixed cells, whole cells independent of sizing and morphology, as well as of nuclei. We applied this method to dermal fibroblasts derived from six patients with different segmental progeria syndromes and defined phenotype associated pathway signatures with variant associated expression modifiers. These results validate the applicability of our method to highlight genotype-expression relationships for molecular phenotyping of individual cells derived from human patients.https://doi.org/10.1038/s41598-022-07874-1 |
| spellingShingle | Orr Shomroni Maren Sitte Julia Schmidt Sabnam Parbin Fabian Ludewig Gökhan Yigit Laura Cecilia Zelarayan Katrin Streckfuss-Bömeke Bernd Wollnik Gabriela Salinas A novel single-cell RNA-sequencing approach and its applicability connecting genotype to phenotype in ageing disease Scientific Reports |
| title | A novel single-cell RNA-sequencing approach and its applicability connecting genotype to phenotype in ageing disease |
| title_full | A novel single-cell RNA-sequencing approach and its applicability connecting genotype to phenotype in ageing disease |
| title_fullStr | A novel single-cell RNA-sequencing approach and its applicability connecting genotype to phenotype in ageing disease |
| title_full_unstemmed | A novel single-cell RNA-sequencing approach and its applicability connecting genotype to phenotype in ageing disease |
| title_short | A novel single-cell RNA-sequencing approach and its applicability connecting genotype to phenotype in ageing disease |
| title_sort | novel single cell rna sequencing approach and its applicability connecting genotype to phenotype in ageing disease |
| url | https://doi.org/10.1038/s41598-022-07874-1 |
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