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Nucleic Acids Res. 2014;42(16):e127. doi: 10.1093/nar/gku604. Epub 2014 Jul 10.

In vivo generation of DNA sequence diversity for cellular barcoding.

Author information

1
Watson School of Biological Sciences, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
2
Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
3
Watson School of Biological Sciences, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA zador@cshl.edu.

Abstract

Heterogeneity is a ubiquitous feature of biological systems. A complete understanding of such systems requires a method for uniquely identifying and tracking individual components and their interactions with each other. We have developed a novel method of uniquely tagging individual cells in vivo with a genetic 'barcode' that can be recovered by DNA sequencing. Our method is a two-component system comprised of a genetic barcode cassette whose fragments are shuffled by Rci, a site-specific DNA invertase. The system is highly scalable, with the potential to generate theoretical diversities in the billions. We demonstrate the feasibility of this technique in Escherichia coli. Currently, this method could be employed to track the dynamics of populations of microbes through various bottlenecks. Advances of this method should prove useful in tracking interactions of cells within a network, and/or heterogeneity within complex biological samples.

PMID:
25013177
PMCID:
PMC4176322
DOI:
10.1093/nar/gku604
[Indexed for MEDLINE]
Free PMC Article

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