Multiplexing clonality: combining RGB marking and genetic barcoding
RGB marking and DNA barcoding are two cutting-edge technologies in the field of clonal cell marking. To combine the virtues of both approaches, we equipped LeGO vectors encoding red, green or blue fluorescent proteins with complex DNA barcodes carrying color-specific signatures. For these vectors, w...
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| Format: | Online |
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Oxford University Press
2014
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| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3985654/ |
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pubmed-39856542014-04-18 Multiplexing clonality: combining RGB marking and genetic barcoding Cornils, Kerstin Thielecke, Lars Hüser, Svenja Forgber, Michael Thomaschewski, Michael Kleist, Nadja Hussein, Kais Riecken, Kristoffer Volz, Tassilo Gerdes, Sebastian Glauche, Ingmar Dahl, Andreas Dandri, Maura Roeder, Ingo Fehse, Boris Methods Online RGB marking and DNA barcoding are two cutting-edge technologies in the field of clonal cell marking. To combine the virtues of both approaches, we equipped LeGO vectors encoding red, green or blue fluorescent proteins with complex DNA barcodes carrying color-specific signatures. For these vectors, we generated highly complex plasmid libraries that were used for the production of barcoded lentiviral vector particles. In proof-of-principle experiments, we used barcoded vectors for RGB marking of cell lines and primary murine hepatocytes. We applied single-cell polymerase chain reaction to decipher barcode signatures of individual RGB-marked cells expressing defined color hues. This enabled us to prove clonal identity of cells with one and the same RGB color. Also, we made use of barcoded vectors to investigate clonal development of leukemia induced by ectopic oncogene expression in murine hematopoietic cells. In conclusion, by combining RGB marking and DNA barcoding, we have established a novel technique for the unambiguous genetic marking of individual cells in the context of normal regeneration as well as malignant outgrowth. Moreover, the introduction of color-specific signatures in barcodes will facilitate studies on the impact of different variables (e.g. vector type, transgenes, culture conditions) in the context of competitive repopulation studies. Oxford University Press 2014-04 2014-01-28 /pmc/articles/PMC3985654/ /pubmed/24476916 http://dx.doi.org/10.1093/nar/gku081 Text en © The Author(s) 2014. Published by Oxford University Press. http://creativecommons.org/licenses/by/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| author |
Cornils, Kerstin Thielecke, Lars Hüser, Svenja Forgber, Michael Thomaschewski, Michael Kleist, Nadja Hussein, Kais Riecken, Kristoffer Volz, Tassilo Gerdes, Sebastian Glauche, Ingmar Dahl, Andreas Dandri, Maura Roeder, Ingo Fehse, Boris |
| spellingShingle |
Cornils, Kerstin Thielecke, Lars Hüser, Svenja Forgber, Michael Thomaschewski, Michael Kleist, Nadja Hussein, Kais Riecken, Kristoffer Volz, Tassilo Gerdes, Sebastian Glauche, Ingmar Dahl, Andreas Dandri, Maura Roeder, Ingo Fehse, Boris Multiplexing clonality: combining RGB marking and genetic barcoding |
| author_facet |
Cornils, Kerstin Thielecke, Lars Hüser, Svenja Forgber, Michael Thomaschewski, Michael Kleist, Nadja Hussein, Kais Riecken, Kristoffer Volz, Tassilo Gerdes, Sebastian Glauche, Ingmar Dahl, Andreas Dandri, Maura Roeder, Ingo Fehse, Boris |
| author_sort |
Cornils, Kerstin |
| title |
Multiplexing clonality: combining RGB marking and genetic barcoding |
| title_short |
Multiplexing clonality: combining RGB marking and genetic barcoding |
| title_full |
Multiplexing clonality: combining RGB marking and genetic barcoding |
| title_fullStr |
Multiplexing clonality: combining RGB marking and genetic barcoding |
| title_full_unstemmed |
Multiplexing clonality: combining RGB marking and genetic barcoding |
| title_sort |
multiplexing clonality: combining rgb marking and genetic barcoding |
| description |
RGB marking and DNA barcoding are two cutting-edge technologies in the field of clonal cell marking. To combine the virtues of both approaches, we equipped LeGO vectors encoding red, green or blue fluorescent proteins with complex DNA barcodes carrying color-specific signatures. For these vectors, we generated highly complex plasmid libraries that were used for the production of barcoded lentiviral vector particles. In proof-of-principle experiments, we used barcoded vectors for RGB marking of cell lines and primary murine hepatocytes. We applied single-cell polymerase chain reaction to decipher barcode signatures of individual RGB-marked cells expressing defined color hues. This enabled us to prove clonal identity of cells with one and the same RGB color. Also, we made use of barcoded vectors to investigate clonal development of leukemia induced by ectopic oncogene expression in murine hematopoietic cells. In conclusion, by combining RGB marking and DNA barcoding, we have established a novel technique for the unambiguous genetic marking of individual cells in the context of normal regeneration as well as malignant outgrowth. Moreover, the introduction of color-specific signatures in barcodes will facilitate studies on the impact of different variables (e.g. vector type, transgenes, culture conditions) in the context of competitive repopulation studies. |
| publisher |
Oxford University Press |
| publishDate |
2014 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3985654/ |
| _version_ |
1612077712241852416 |