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dc.contributor.authorMarshall, Ryan
dc.contributor.authorBeisel, Chase L
dc.contributor.authorNoireaux, Vincent
dc.date.accessioned2020-11-09T14:19:21Z
dc.date.available2020-11-09T14:19:21Z
dc.date.issued2020-06-03
dc.identifier.citationSTAR Protoc. 2020 Jun 3;1(1):100003. doi: 10.1016/j.xpro.2019.100003.en_US
dc.identifier.pmid33111065
dc.identifier.doi10.1016/j.xpro.2019.100003
dc.identifier.urihttp://hdl.handle.net/10033/622564
dc.description.abstractWe present a protocol to rapidly test DNA binding and cleavage activity by CRISPR nucleases using cell-free transcription-translation (TXTL). Nuclease activity is assessed by adding DNA encoding a nuclease, a guide RNA, and a targeted reporter to a TXTL reaction and by measuring the fluorescence for several h. The reactions, performed in a few microliters, allow for parallel testing of many nucleases and guide RNAs. The protocol includes representative results for (d)Cas9 from Streptococcus pyogenes targeting a GFP reporter gene. For complete information on the generation and use of this protocol, please refer to the paper by Marshall et al. (2018).en_US
dc.language.isoenen_US
dc.publisherElsevier (CellPress)en_US
dc.rightsAttribution-NonCommercial-ShareAlike 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.titleRapid Testing of CRISPR Nucleases and Guide RNAs in an Cell-Free Transcription-Translation System.en_US
dc.typeArticleen_US
dc.identifier.eissn2666-1667
dc.contributor.departmentHIRI, Helmholtz-Institut für RNA-basierte Infektionsforschung, Josef-Shneider Strasse 2, 97080 Würzburg, Germany.en_US
dc.identifier.journalSTAR protocolsen_US
dc.source.volume1
dc.source.issue1
dc.source.beginpage100003
dc.source.endpage
refterms.dateFOA2020-11-09T14:19:23Z
dc.source.journaltitleSTAR protocols
dc.source.countryUnited States


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Attribution-NonCommercial-ShareAlike 4.0 International
Except where otherwise noted, this item's license is described as Attribution-NonCommercial-ShareAlike 4.0 International