Expanded hammerhead ribozymes containing addressable three-way junctions

Wieland, Markus; Gfell, Manuela; Hartig, Jörg S.

Expanded hammerhead ribozymes containing addressable three-way junctions

Dateien

wieland_opus_117254.pdfGröße: 819.79 KBDownloads: 224

Datum

2009

Autor:innen

Wieland, Markus

Gfell, Manuela

Hartig, Jörg S.

Publikationstyp

Zeitschriftenartikel

unikn.publication.listelement.citation.prefix.version.undefined

RNA. 2009, 15(5), pp. 968-976. ISSN 1355-8382. eISSN 1469-9001. Available under: doi: 10.1261/rna.1220309

Zusammenfassung

Recently, hammerhead ribozyme (HHR) motifs have been utilized as powerful tools for gene regulation. Here we present a novel design of expanded full-length HHRs that allows attaching additional functionalities to the ribozyme. These features allowed us to construct a very efficient artificial riboswitch in bacteria. Following the design of naturally occurring three-way junctions we attached an additional helix (IV) to stem I of the HHR while maintaining very fast cleavage rates. We found that the cleavage activity strongly depends on the exact design of the junction site. Incorporation of the novel ribozyme scaffold into a bacterial mRNA allowed the control of gene expression mediated by autocatalytic cleavage of the ribozyme. Appending an aptamer to the newly introduced stem enabled the identification of very powerful theophylline-inducible RNA switches by in vivo screening. Further investigations revealed a cascading system operating beyond the ribozyme-dependent mechanism. In conclusion, we extended the hammerhead toolbox for synthetic biology applications by providing an additional position for the attachment of regulatory modules for in vivo control of gene expression.

Fachgebiet (DDC)

540 Chemie

Zitieren

ISO 690

WIELAND, Markus, Manuela GFELL, Jörg S. HARTIG, 2009. Expanded hammerhead ribozymes containing addressable three-way junctions. In: RNA. 2009, 15(5), pp. 968-976. ISSN 1355-8382. eISSN 1469-9001. Available under: doi: 10.1261/rna.1220309

BibTex

@article{Wieland2009-05Expan-1011,
  year={2009},
  doi={10.1261/rna.1220309},
  title={Expanded hammerhead ribozymes containing addressable three-way junctions},
  number={5},
  volume={15},
  issn={1355-8382},
  journal={RNA},
  pages={968--976},
  author={Wieland, Markus and Gfell, Manuela and Hartig, Jörg S.}
}

RDF

<rdf:RDF
    xmlns:dcterms="http://purl.org/dc/terms/"
    xmlns:dc="http://purl.org/dc/elements/1.1/"
    xmlns:rdf="http://www.w3.org/1999/02/22-rdf-syntax-ns#"
    xmlns:bibo="http://purl.org/ontology/bibo/"
    xmlns:dspace="http://digital-repositories.org/ontologies/dspace/0.1.0#"
    xmlns:foaf="http://xmlns.com/foaf/0.1/"
    xmlns:void="http://rdfs.org/ns/void#"
    xmlns:xsd="http://www.w3.org/2001/XMLSchema#" > 
  <rdf:Description rdf:about="https://kops.uni-konstanz.de/server/rdf/resource/123456789/1011">
    <foaf:homepage rdf:resource="http://localhost:8080/"/>
    <void:sparqlEndpoint rdf:resource="http://localhost/fuseki/dspace/sparql"/>
    <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/29"/>
    <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/52"/>
    <dcterms:hasPart rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/1011/1/wieland_opus_117254.pdf"/>
    <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/29"/>
    <dc:language>eng</dc:language>
    <dcterms:bibliographicCitation>RNA ; 15 (2009), 5. - S. 968-976</dcterms:bibliographicCitation>
    <dspace:hasBitstream rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/1011/1/wieland_opus_117254.pdf"/>
    <bibo:uri rdf:resource="http://kops.uni-konstanz.de/handle/123456789/1011"/>
    <dcterms:title>Expanded hammerhead ribozymes containing addressable three-way junctions</dcterms:title>
    <dc:contributor>Wieland, Markus</dc:contributor>
    <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-03-22T17:54:42Z</dc:date>
    <dc:creator>Hartig, Jörg S.</dc:creator>
    <dc:contributor>Gfell, Manuela</dc:contributor>
    <dcterms:rights rdf:resource="https://rightsstatements.org/page/InC/1.0/"/>
    <dc:creator>Wieland, Markus</dc:creator>
    <dcterms:abstract>Recently, hammerhead ribozyme (HHR) motifs have been utilized as powerful tools for gene regulation. Here we present a novel design of expanded full-length HHRs that allows attaching additional functionalities to the ribozyme. These features allowed us to construct a very efficient artificial riboswitch in bacteria. Following the design of naturally occurring three-way junctions we attached an additional helix (IV) to stem I of the HHR while maintaining very fast cleavage rates. We found that the cleavage activity strongly depends on the exact design of the junction site. Incorporation of the novel ribozyme scaffold into a bacterial mRNA allowed the control of gene expression mediated by autocatalytic cleavage of the ribozyme. Appending an aptamer to the newly introduced stem enabled the identification of very powerful theophylline-inducible RNA switches by in vivo screening. Further investigations revealed a cascading system operating beyond the ribozyme-dependent mechanism. In conclusion, we extended the hammerhead toolbox for synthetic biology applications by providing an additional position for the attachment of regulatory modules for in vivo control of gene expression.</dcterms:abstract>
    <dc:contributor>Hartig, Jörg S.</dc:contributor>
    <dc:rights>terms-of-use</dc:rights>
    <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/52"/>
    <dc:creator>Gfell, Manuela</dc:creator>
    <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-03-22T17:54:42Z</dcterms:available>
    <dcterms:issued>2009-05</dcterms:issued>
  </rdf:Description>
</rdf:RDF>

Universitätsbibliographie

Ja