English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
Add to Basket
  Local TagsRelease HistoryDetailsSummary

Released
Conference Paper

Live-cell single-particle tracking photoactivated localization microscopy of Cascade-mediated DNA surveillance

MPS-Authors
/persons/resource/persons254776

Turkowyd,  Bartosz
Department of Systems and Synthetic Microbiology, Max Planck Institute for Terrestrial Microbiology, Max Planck Society;

/persons/resource/persons254561

Müller-Esparza,  Hanna
Max Planck Research Group Prokaryotic small RNA Biology, Alumni, Max Planck Institute for Terrestrial Microbiology, Max Planck Society;

Climenti,  Vanessa
Max Planck Research Group Prokaryotic small RNA Biology, Alumni, Max Planck Institute for Terrestrial Microbiology, Max Planck Society;

Steube,  Niklas
Department of Systems and Synthetic Microbiology, Max Planck Institute for Terrestrial Microbiology, Max Planck Society;

/persons/resource/persons256432

Endesfelder,  Ulrike
Department of Systems and Synthetic Microbiology, Max Planck Institute for Terrestrial Microbiology, Max Planck Society;

/persons/resource/persons254626

Randau,  Lennart
Max Planck Research Group Prokaryotic small RNA Biology, Alumni, Max Planck Institute for Terrestrial Microbiology, Max Planck Society;

External Resource
No external resources are shared
Fulltext (restricted access)
There are currently no full texts shared for your IP range.
Fulltext (public)
There are no public fulltexts stored in PuRe
Supplementary Material (public)
There is no public supplementary material available
Citation

Turkowyd, B., Müller-Esparza, H., Climenti, V., Steube, N., Endesfelder, U., & Randau, L. (2019). Live-cell single-particle tracking photoactivated localization microscopy of Cascade-mediated DNA surveillance. In CRISPR-CAS ENZYMES (pp. 133-171).


Cite as: https://hdl.handle.net/21.11116/0000-0008-BF60-5
Abstract
Type I CRISPR-Cas systems utilize small CRISPR RNA (crRNA) molecules to
scan DNA strands for target regions. Different crRNAs are bound by
several CRISPR-associated (Cas) protein subunits that form the stable
ribonucleoprotein complex Cascade. The Cascade-mediated DNA surveillance
process requires a sufficient degree of base-complementarity between
crRNA and target sequences and relies on the recognition of small DNA
motifs, termed protospacer adjacent motifs. Recently, superresolution
microscopy and single-particle tracking methods have been developed to
follow individual protein complexes in live cells. Here, we described
how this technology can be adapted to visualize the DNA scanning process
of Cascade assemblies in Escherichia coli cells. The activity of
recombinant Type I-Fv Cascade complexes of Shewanella putrefaciens CN-32
serves as a model system that facilitates comparative studies for many
of the diverse CRISPR-Cas systems.