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Cellular barcoding tool for clonal analysis in the hematopoietic system.
| Content Provider | Semantic Scholar |
|---|---|
| Author | Gerrits, Alice Dykstra, Brad Kalmykowa, O. J. Klauke, Karin Verovskaya, Evgenia Broekhuis, M. J. C. Haan, Gerald De Bystrykh, L. V. |
| Copyright Year | 2010 |
| Abstract | Clonal analysis is important for many areas of hematopoietic stem cell research, including in vitro cell expansion, gene therapy, and cancer progression and treatment. A common approach to measure clonality of retrovirally transduced cells is to perform integration site analysis using Southern blotting or polymerase chain reaction-based methods. Although these methods are useful in principle, they generally provide a low-resolution, biased, and incomplete assessment of clonality. To overcome those limitations, we labeled retroviral vectors with random sequence tags or "barcodes." On integration, each vector introduces a unique, identifiable, and heritable mark into the host cell genome, allowing the clonal progeny of each cell to be tracked over time. By coupling the barcoding method to a sequencing-based detection system, we could identify major and minor clones in 2 distinct cell culture systems in vitro and in a long-term transplantation setting. In addition, we demonstrate how clonal analysis can be complemented with transgene expression and integration site analysis. This cellular barcoding tool permits a simple, sensitive assessment of clonality and holds great promise for future gene therapy protocols in humans, and any other applications when clonal tracking is important. |
| File Format | PDF HTM / HTML |
| DOI | 10.1182/blood-2009-06-229757 |
| PubMed reference number | 20093403 |
| Journal | Medline |
| Volume Number | 115 |
| Issue Number | 13 |
| Alternate Webpage(s) | http://eriba.umcg.nl/wp-content/uploads/2016/01/DeHaan-2010-01-21-Blood.pdf |
| Alternate Webpage(s) | https://doi.org/10.1182/blood-2009-06-229757 |
| Journal | Blood |
| Language | English |
| Access Restriction | Open |
| Content Type | Text |
| Resource Type | Article |
