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 language | English |
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Patent number | WO2019072596 |
Priority date | 10/10/17 |
Publication status | Published - 18 Apr 2019 |