Your browser doesn't support javascript.
loading
Nuclease dead Cas9 is a programmable roadblock for DNA replication.
Whinn, Kelsey S; Kaur, Gurleen; Lewis, Jacob S; Schauer, Grant D; Mueller, Stefan H; Jergic, Slobodan; Maynard, Hamish; Gan, Zhong Yan; Naganbabu, Matharishwan; Bruchez, Marcel P; O'Donnell, Michael E; Dixon, Nicholas E; van Oijen, Antoine M; Ghodke, Harshad.
Afiliação
  • Whinn KS; School of Chemistry and Molecular Bioscience and Molecular Horizons, University of Wollongong, Wollongong, New South Wales, 2522, Australia.
  • Kaur G; Illawarra Health and Medical Research Institute, Wollongong, New South Wales, 2522, Australia.
  • Lewis JS; School of Chemistry and Molecular Bioscience and Molecular Horizons, University of Wollongong, Wollongong, New South Wales, 2522, Australia.
  • Schauer GD; Illawarra Health and Medical Research Institute, Wollongong, New South Wales, 2522, Australia.
  • Mueller SH; School of Chemistry and Molecular Bioscience and Molecular Horizons, University of Wollongong, Wollongong, New South Wales, 2522, Australia.
  • Jergic S; Illawarra Health and Medical Research Institute, Wollongong, New South Wales, 2522, Australia.
  • Maynard H; Howard Hughes Medical Institute, Rockefeller University, New York, NY, 10065, USA.
  • Gan ZY; School of Chemistry and Molecular Bioscience and Molecular Horizons, University of Wollongong, Wollongong, New South Wales, 2522, Australia.
  • Naganbabu M; Illawarra Health and Medical Research Institute, Wollongong, New South Wales, 2522, Australia.
  • Bruchez MP; School of Chemistry and Molecular Bioscience and Molecular Horizons, University of Wollongong, Wollongong, New South Wales, 2522, Australia.
  • O'Donnell ME; Illawarra Health and Medical Research Institute, Wollongong, New South Wales, 2522, Australia.
  • Dixon NE; School of Chemistry and Molecular Bioscience and Molecular Horizons, University of Wollongong, Wollongong, New South Wales, 2522, Australia.
  • van Oijen AM; School of Chemistry and Molecular Bioscience and Molecular Horizons, University of Wollongong, Wollongong, New South Wales, 2522, Australia.
  • Ghodke H; Department of Chemistry and Molecular Biosensors and Imaging Center, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, Pennsylvania, 15213, USA.
Sci Rep ; 9(1): 13292, 2019 09 16.
Article em En | MEDLINE | ID: mdl-31527759
ABSTRACT
Limited experimental tools are available to study the consequences of collisions between DNA-bound molecular machines. Here, we repurpose a catalytically inactivated Cas9 (dCas9) construct as a generic, novel, targetable protein-DNA roadblock for studying mechanisms underlying enzymatic activities on DNA substrates in vitro. We illustrate the broad utility of this tool by demonstrating replication fork arrest by the specifically bound dCas9-guideRNA complex to arrest viral, bacterial and eukaryotic replication forks in vitro.
Assuntos
Texto completo
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: DNA Bacteriano / RNA Guia de Cinetoplastídeos / Replicação do DNA / Escherichia coli / Proteína 9 Associada à CRISPR Idioma: En Ano de publicação: 2019 Tipo de documento: Article
Texto completo
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: DNA Bacteriano / RNA Guia de Cinetoplastídeos / Replicação do DNA / Escherichia coli / Proteína 9 Associada à CRISPR Idioma: En Ano de publicação: 2019 Tipo de documento: Article