Thermostable Cas9 nucleases with reduced off-target activity

J. van der Oost (Inventor), R. van Kranenburg (Inventor), E.F. Bosma (Inventor), I. Mougiakos (Inventor), P. Mohanraju (Inventor)

Research output: PatentOther research output

Abstract

ThermoCas9 is identified and characterized from the thermophilic bacterium Geobacillus thermodenitrificans T12. Experiments show how in vitro ThermoCas9 is active between 20 and 70 °C, has stringent PAM-preference at lower temperatures, tolerates fewer spacer-protospacer mismatches than SpCas9 and its activity at elevated temperatures depends on the sgRNA-structure. Described are ThermoCas9-based engineering tools for gene deletion and transcriptional silencing at 5 °C in Bacillus smithii and for gene deletion at 37 °C in Pseudomonas putida
Original languageEnglish
Patent numberWO2019072596
Priority date10/10/17
Publication statusPublished - 18 Apr 2019

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