17
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: not found
      • Article: not found

      Multidimensional Control of Cas9 by Evolved RNA Polymerase-Based Biosensors

      1 , 1 , 1
      ACS Chemical Biology
      American Chemical Society (ACS)

      Read this article at

      ScienceOpenPublisherPMC
      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Abstract

          Systems to control Cas9 with spatial and temporal precision offer opportunities to decrease side effects, protect sensitive tissues, and create gene therapies that are only activated at defined times and places. Here, we present the design of new Cas9 controllers based on RNA polymerase (RNAP)-based biosensors that produce gRNAs, thereby regulating target knockout. After development and validation of a new abscisic acid-inducible biosensor to control Cas9, we lowered the background of the system using continuous evolution. To showcase the versatility of the approach, we designed biosensors that measure medically relevant protein-protein interactions to drive knockout. Finally, to test whether orthogonal RNAP biosensors could integrate multiple input signals to drive multiple gRNA-based outputs with a single Cas9 protein, we designed an "on-switch/off switch" controller. The addition of one input activates the "on switch" and induces knockout, while the addition of a second input activates the "off switch" and produces a gRNA that directs the Cas9 protein to degrade the "on switch" gRNA vector, thereby deactivating it. This combined activation and deactivation system displayed very low background and inducible target knockout using different combinations of small-molecule treatment. Our results establish engineered RNAP biosensors as deployable Cas9 control elements and open up new opportunities for driving genetic editing technologies by diverse input signals.

          Related collections

          Author and article information

          Journal
          ACS Chemical Biology
          ACS Chem. Biol.
          American Chemical Society (ACS)
          1554-8929
          1554-8937
          September 15 2017
          February 16 2018
          August 22 2017
          February 16 2018
          : 13
          : 2
          : 431-437
          Affiliations
          [1 ]Department of Chemistry, The University of Chicago, Chicago, Illinois 60637, United States
          Article
          10.1021/acschembio.7b00532
          5828011
          28809467
          7a375ba0-f288-4647-84b0-c3d7789a95ab
          © 2018
          History

          Comments

          Comment on this article

          scite_
          0
          0
          0
          0
          Smart Citations
          0
          0
          0
          0
          Citing PublicationsSupportingMentioningContrasting
          View Citations

          See how this article has been cited at scite.ai

          scite shows how a scientific paper has been cited by providing the context of the citation, a classification describing whether it supports, mentions, or contrasts the cited claim, and a label indicating in which section the citation was made.

          Similar content4,355

          Cited by12