Genetic Barcodes Facilitate Competitive Clonal Analyses In Vivo

Research output: Contribution to journalResearch articleContributedpeer-review



Monitoring the fate of individual cell clones is an important task to better understand normal tissue regeneration, for example after hematopoietic stem cell (HSC) transplantation, but also cancerogenesis. Based on their integration into the host cell's genome, retroviral vectors are commonly used to stably mark target cells and their progeny. The development of genetic barcoding techniques has opened new possibilities to determine clonal composition and dynamics in great detail. A modular genetic barcode was recently introduced consisting of 32 variable positions (BC32) with a customized backbone, and its advantages were demonstrated with regard to barcode calling and quantification. The study presented applied the BC32 system in a complex in vivo situation, namely to analyze clonal reconstitution dynamics for HSC grafts consisting of up to three cell populations with distinguishable barcodes using different alpha- and lentiviral vectors. In a competitive transplantation setup, it was possible to follow the differently marked cell populations within individual animals. This enabled the clonal contribution of the different BC32 constructs during reconstitution and long-term hematopoiesis in the peripheral blood and the spatial distribution in bone marrow and spleen to be identified. Thus, it was demonstrated that the system allows the output of individually marked cells to be tracked in vivo and their influence on clonal dynamics to be analyzed. Successful application of the BC32 system in a complex, competitive in vivo situation provided proof-of-principle that its high complexity and the large Hamming distance between individual barcodes, combined with the easy customization, facilitate efficient and precise quantification, even without prior knowledge of individual barcode sequences. Importantly, simultaneous high-sensitivity analyses of different cell populations in single animals may significantly reduce numbers of animals required to investigate specific scientific questions in accordance with RRR principles. It is concluded that this BC32 system will be excellently suited for various research applications in regenerative medicine and cancer biology.


Original languageEnglish
Pages (from-to)926-937
Number of pages12
JournalHuman Gene Therapy
Issue number10
Publication statusPublished - Oct 2017

External IDs

Scopus 85030328764
PubMed 28847169
ORCID /0000-0002-2524-1199/work/142251490
ORCID /0000-0002-5726-4491/work/153109649


Sustainable Development Goals


  • Animals, Bone Marrow Cells/cytology, Clonal Evolution/genetics, Clone Cells, Gene Expression, Gene Order, Genetic Vectors/genetics, Hematopoietic Stem Cells/cytology, Humans, Lentivirus/genetics, Mice, Promoter Regions, Genetic, Transduction, Genetic, Transgenes