Abstract
Mutational signatures have been identified in cancer genomes, providing information about the causes of cancer and treatment vulnerabilities. This protocol describes an assay to determine the genotoxic mechanisms underlying these signatures using cord-blood derived hematopoietic stem and progenitor cells (CB-HSPCs). CB-HSPCs have a low mutation background, enabling sensitive detection of mutations. First, CB-HSPCs are exposed in vitro, sorted, and clonally expanded. This expansion enables whole-genome sequencing to detect the mutation load and respective patterns induced during genotoxic exposure. For complete details on the use and execution of this protocol, please refer to de Kanter et al. (2021).
| Original language | English |
|---|---|
| Article number | 101361 |
| Journal | STAR Protocols |
| Volume | 3 |
| Issue number | 2 |
| Early online date | 6 May 2022 |
| DOIs | |
| Publication status | Published - 17 Jun 2022 |
Bibliographical note
Funding Information:This research was supported by funding from a NWO Vidi grant to R.v.B, no. 016.Vidi.171.023, and the Oncode Institute . The authors want to thank M.A. Satzl for providing pictures of the CMBC isolation.
Funding Information:
This research was supported by funding from a NWO Vidi grant to R.v.B, no. 016.Vidi.171.023, and the Oncode Institute. The authors want to thank M.A. Satzl for providing pictures of the CMBC isolation. A.R.H. and A.J.C.N.v.L. performed the experiments. A.R.H. performed the bioinformatic analyses. A.J.C.N.v.L, A.R.H. F.P, E.J.M.B, and J.K.d.K. wrote the manuscript. All authors read, revised, and approved the manuscript. A.R.H. A.v.L. and R.v.B. are named as inventors on a patent application filed resulting from this work.
Publisher Copyright:
© 2022 The Author(s)
