Site-specific recombinatorics: in situ cellular barcoding with the Cre Lox system
Details
Publication Year 2016,Volume 10,Issue #1,Page 43
Journal Title
BMC Syst Biol
Publication Type
Journal Article
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.
Publisher
BioMed Central
Research Division(s)
Cancer And Haematology; Molecular Medicine
PubMed ID
27363727
NHMRC Grants
NHMRC/1052195
Terms of Use/Rights Notice
Refer to copyright notice on published article.


Creation Date: 2016-08-09 12:01:43
Last Modified: 2016-08-10 09:42:17
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