Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Fluorescent CRISPR Adaptation Reporter for rapid quantification of spacer acquisition
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Microbiology.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Microbiology.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Microbiology.
Show others and affiliations
2017 (English)In: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 7, article id 10392Article in journal (Refereed) Published
Abstract [en]

CRISPR-Cas systems are adaptive prokaryotic immune systems protecting against horizontally transferred DNA or RNA such as viruses and other mobile genetic elements. Memory of past invaders is stored as spacers in CRISPR loci in a process called adaptation. Here we developed a novel assay where spacer integration results in fluorescence, enabling detection of memory formation in single cells and quantification of as few as 0.05% cells with expanded CRISPR arrays in a bacterial population. Using this fluorescent CRISPR Adaptation Reporter (f-CAR), we quantified adaptation of the two CRISPR arrays of the type I-E CRISPR-Cas system in Escherichia coli, and confirmed that more integration events are targeted to CRISPR-II than to CRISPR-I. The f-CAR conveniently analyzes and compares many samples, allowing new insights into adaptation. For instance, we show that in an E. coli culture the majority of acquisition events occur in late exponential phase.

Place, publisher, year, edition, pages
2017. Vol. 7, article id 10392
Keywords [en]
quorum sensing controls, escherichia-coli, cas adaptation, immune-system, host factor, dna, resistance, elements, repeats, bacteriophage
National Category
Microbiology
Identifiers
URN: urn:nbn:se:uu:diva-334399DOI: 10.1038/s41598-017-10876-zISI: 000408997700091OAI: oai:DiVA.org:uu-334399DiVA, id: diva2:1163840
Available from: 2017-12-08 Created: 2017-12-08 Last updated: 2017-12-08Bibliographically approved

Open Access in DiVA

fulltext(3335 kB)38 downloads
File information
File name FULLTEXT01.pdfFile size 3335 kBChecksum SHA-512
52ba9bbc59bd8310d1cf721364c7b92b0b086070ae1cf3ab11f354df9f17945033fdd890badc5f6c618ac8fd6976340bbc2ce9ef13bf621b3958cc3e53130909
Type fulltextMimetype application/pdf

Other links

Publisher's full text

Search in DiVA

By author/editor
Amlinger, LinaHoekzema, MirtheWagner, Gerhart E. H.Koskiniemi, SannaLundgren, Magnus
By organisation
MicrobiologyDepartment of Cell and Molecular Biology
In the same journal
Scientific Reports
Microbiology

Search outside of DiVA

GoogleGoogle Scholar
Total: 38 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

doi
urn-nbn

Altmetric score

doi
urn-nbn
Total: 145 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf