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  • 标题:Nuclease dead Cas9 is a programmable roadblock for DNA replication
  • 本地全文:下载
  • 作者:Kelsey S. Whinn ; Gurleen Kaur ; Jacob S. Lewis
  • 期刊名称:Scientific Reports
  • 电子版ISSN:2045-2322
  • 出版年度:2019
  • 卷号:9
  • 期号:1
  • 页码:1-9
  • DOI:10.1038/s41598-019-49837-z
  • 出版社:Springer Nature
  • 摘要: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.
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