Site-specific recombinatorics: in situ cellular barcoding with the Cre Lox system

Tom S. Weber, Mark Dukes, Denise C. Miles, Stefan P. Glaser, Shalin H. Naik, Ken R. Duffy

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Background: Cellular barcoding is a recently developed biotechnology tool that enables the familial identification of progeny of individual cells in vivo. In immunology, it has been used to track the burst-sizes of multiple distinct responding T cells over several adaptive immune responses. In the study of hematopoiesis, it revealed fate heterogeneity amongst phenotypically identical multipotent cells. Most existing approaches rely on ex vivo viral transduction of cells with barcodes followed by adoptive transfer into an animal, which works well for some systems, but precludes barcoding cells in their native environment such as those inside solid tissues.
Results: With a view to overcoming this limitation, we propose a new design for a genetic barcoding construct based on the Cre Lox system that induces randomly created stable barcodes in cells in situ by exploiting inherent sequence distance constraints during site-specific recombination. We identify the cassette whose provably maximal code diversity is several orders of magnitude higher than what is attainable with previously considered Cre Lox barcoding approaches, exceeding the number of lymphocytes or hematopoietic progenitor cells in mice.
Conclusions: Its high diversity and in situ applicability, make the proposed Cre Lox based tagging system suitable for whole tissue or even whole animal barcoding. Moreover, it can be built using established technology.
LanguageEnglish
Article number43
Number of pages13
JournalBMC Systems Biology
Volume10
DOIs
Publication statusPublished - 30 Jun 2016

Fingerprint

Animals
Immunology
Tissue
T-cells
Lymphocytes
Cell
Biotechnology
Cells
Hematopoiesis
Adoptive Transfer
Lymphocyte Count
Adaptive Immunity
Hematopoietic Stem Cells
Allergy and Immunology
Immune Response
Genetic Recombination
Tagging
Burst
Recombination
Mouse

Keywords

  • cell fate tracking
  • cellular barcoding
  • combinatorial explosion
  • Cre Lox system
  • DNA stochastic programme

Cite this

Weber, T. S., Dukes, M., Miles, D. C., Glaser, S. P., Naik, S. H., & Duffy, K. R. (2016). Site-specific recombinatorics: in situ cellular barcoding with the Cre Lox system. BMC Systems Biology, 10, [43]. https://doi.org/10.1186/s12918-016-0290-3
Weber, Tom S. ; Dukes, Mark ; Miles, Denise C. ; Glaser, Stefan P. ; Naik, Shalin H. ; Duffy, Ken R. / Site-specific recombinatorics : in situ cellular barcoding with the Cre Lox system. In: BMC Systems Biology. 2016 ; Vol. 10.
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Site-specific recombinatorics : in situ cellular barcoding with the Cre Lox system. / Weber, Tom S.; Dukes, Mark; Miles, Denise C.; Glaser, Stefan P.; Naik, Shalin H.; Duffy, Ken R.

In: BMC Systems Biology, Vol. 10, 43, 30.06.2016.

Research output: Contribution to journalArticle

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