Sangam: A Confluence of Knowledge Streams

Lentiviral and targeted cellular barcoding reveals ongoing clonal dynamics of cell lines in vitro and in vivo

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dc.contributor Biological Sciences
dc.contributor Fralin Life Sciences Institute
dc.creator Porter, Shaina N.
dc.creator Baker, Lee C.
dc.creator Mittelman, David
dc.creator Porteus, Matthew H.
dc.date 2015-07-31T16:40:19Z
dc.date 2015-07-31T16:40:19Z
dc.date 2014-05-30
dc.date 2015-07-31T16:40:19Z
dc.date.accessioned 2023-03-01T18:54:35Z
dc.date.available 2023-03-01T18:54:35Z
dc.identifier Genome Biology. 2014 May 30;15(5):R75
dc.identifier http://hdl.handle.net/10919/55017
dc.identifier https://doi.org/10.1186/gb-2014-15-5-r75
dc.identifier.uri http://localhost:8080/xmlui/handle/CUHPOERS/281843
dc.description Background Cell lines are often regarded as clonal, even though this simplifies what is known about mutagenesis, transformation and other processes that destabilize them over time. Monitoring these clonal dynamics is important for multiple areas of biomedical research, including stem cell and cancer biology. Tracking the contributions of individual cells to large populations, however, has been constrained by limitations in sensitivity and complexity. Results We utilize cellular barcoding methods to simultaneously track the clonal contributions of tens of thousands of cells. We demonstrate that even with optimal culturing conditions, common cell lines including HeLa, K562 and HEK-293 T exhibit ongoing clonal dynamics. Starting a population with a single clone diminishes but does not eradicate this phenomenon. Next, we compare lentiviral and zinc-finger nuclease barcode insertion approaches, finding that the zinc-finger nuclease protocol surprisingly results in reduced clonal diversity. We also document the expected reduction in clonal complexity when cells are challenged with genotoxic stress. Finally, we demonstrate that xenografts maintain clonal diversity to a greater extent than in vitro culturing of the human non-small-cell lung cancer cell line HCC827. Conclusions We demonstrate the feasibility of tracking and quantifying the clonal dynamics of entire cell populations within multiple cultured cell lines. Our results suggest that cell heterogeneity should be considered in the design and interpretation of in vitro culture experiments. Aside from clonal cell lines, we propose that cellular barcoding could prove valuable in modeling the clonal behavior of heterogeneous cell populations over time, including tumor populations treated with chemotherapeutic agents.
dc.description Published version
dc.format application/pdf
dc.format application/pdf
dc.language en_US
dc.rights Creative Commons Attribution 4.0 International
dc.rights http://creativecommons.org/licenses/by/4.0/
dc.rights Porter et al.; licensee BioMed Central Ltd.
dc.title Lentiviral and targeted cellular barcoding reveals ongoing clonal dynamics of cell lines in vitro and in vivo
dc.title Genome Biology
dc.type Article - Refereed
dc.type Text


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